text:
Lidstrom: SDS-PAGE
Return: Lidstrom Lab Protocols
Return: official OpenWetWare SDS-PAGE
Contact Janet Matsen with questions, comments, & corrections.
Gel Prep
- Clean cover plate and thicker spacer plate (75 mM gap)
- Soap and Water: scrub with gloved fingers. Ensure all chunks are gone.
- Rinse with water, then ethanol (for fast drying). Prop up vertically against green clamps for stands so they drain and dry.
- Setup one spacer plate and one cover plate in each gel holder
- The cover plate goes on the side of the spacer plate with the spacers in order to create a small gap between the plates
- Make sure the bottom edges are flat after they are clamped down in the stand.
- Make a fresh 10% (w/vol) APS solution. Don't use an "old" solution; the gel won't polymerize. 10% w/v = 0.1 g/mL.
- 1 month old seems to be safe, but up to 3 months may be ok. It depends on how old the stock bottle is.
- If you want to do a test-scale ( < 2 mL) batch in an eppendorf tube, this scaled-down recipe may help. You can mix the ingredients in an eppendorf and see how fast they polymerize, or whether polymerization fails all together.
- Also, if the APS bottle's contents look wet more than powdery, you might not get polymerization at all.
- 1 month old seems to be safe, but up to 3 months may be ok. It depends on how old the stock bottle is.
Pour the Resolving Gel
- Recipe :
- Don't add APS/TEMED until ready to pour
- The APS solution should be < 1 month old. Up to 2 months might be ok, but we don't have great data for this.
- A 3 month old solution failed to cause polymerization & the liquid spilled out from between the plates when released from the stand. -JM 10/2012
- The BioRad protocols insist you should make it fresh every time, but we never do if we have one made within the last 4 weeks
- Use pipette to put gel mix into the gap between the plates. Fill to about the bottom of the green line that is below the short plate.
- leave room for comb + 1 cm of resolving gel (citation: protein gel electrophoresis tips and troubleshooting guide)
- Carefully layer 50%EtOH 50% ddH2O on top of the gel to prevent the top of the gel from drying out
- This also keeps the gel under anaerobic conditions, necessary for polymerization.
- It is very possible to ruin the desired evenness of the resolving gel by squirting this mixture in too fast. Janet has done it!
- Wait 40 minutes for anaerobic polymerization.
- Rinse ethanol/water mix off or you will get bubbles in the gel.
- You can rinse the gel by flushing with tap water from the sink. Afterward, prop the gel against the casting stand on its side so the liquid drains to one edge. Remove this with paper towel.
- Sometimes Janet Matsen skips this step and it seems to work out fine.
- When this is skipped, the gels are drained of the EtOH/H2O mixture, and propped up sideways so the remaining solution pools on one edge. Use a paper towel to suck any remaining liquid up, and repeat again before restoring it to the orientation required for pouring the stacking gel.
- Sometimes Janet Matsen skips this step and it seems to work out fine.
- You can rinse the gel by flushing with tap water from the sink. Afterward, prop the gel against the casting stand on its side so the liquid drains to one edge. Remove this with paper towel.
- Store at 4 oC wrapped in a wet paper towel with either plastic wrap or a plastic bag preventing moisture loss.
- You can also store in the fridge after pouring the stacking gel, and pour the stacking gel later. There doesn't seem to be a benefit to this method, but may allow you to have differences in buffer composition of the resolving and stacking gels. Note, this difference will disappear if you pre-run empty gels as this lab usually does.
Pour the Stacking Gel
- Place gel in gel holder & ensure no water is sitting on top of the resolving gel.
- Remove liquid on top if you didn't already do so. Removal of ethanol/water or butanol is important to prevent bubbles.
- Dry surface of gel carefully with Kimwipe or paper towel
- It can be a little gooey
- Mix components of the amounts in the Gel Mix link. Mix in the order listed.
- recipe (see "stacking gel")
- Don't add APS/TEMED until ready to pour (within a few minutes is ok)
- Use pipette to put gel mix into the gap between the plates.
- Filling to the top reduces the probability of trapping bubbles under the comb when you insert the comb.
- Insert the comb being careful not to trap any bubbles
- It is much easier to avoid bubbles if you fill the space with enough solution that it spills over the top as you put the comb in. It also helps to lower one side completely, then lower the other side down.
- Attach 2 1" wide binder clips to each gel to help hold the comb in while drying. One on either side of the casting stand clamp.
- Leave for 30 minutes while polymerization occurs.
Wrapping up & Storing
- Put binder clips away
- Rinse everything off
- Rinse gel under water with comb in to wash away solution that spilled over.
- Remove comb from gel carefully (don't want to slant the parts that stick up) & soak in soapy water
- Rinse casting stand, & rub the bottom where acrylamide tends to dry on
- rub fingers over the comb to remove crust, rinse, and put on drying rack.
- Wrap gels in a wet paper towel & store in a plastic bag or shrink wrap.
- You can store for a few weeks in the fridge (>= 6 weeks). Leave comb in (fine to remove comb, fill wells with water ALS 10-24-13), and wrap in a wet paper towel and cling wrap or a plastic baggie.
Consolidated/Pictorial Protocol
Sample Prep & Gel Loading
Sample Prep
Prepping from E. coli liquid culture
- If you are boiling cultures and adding total protein, you should know that TB buffer when mixed with loading dye and boiled results in a lot of white precipitate. You can leave the TB in the sample if you want because the precipitate doesn't matter in SDS-PAGE as the gel below shows.
- Experiment that proves TB causes white precipitate:
Does that white precipitate after boiling matter?
- The picture above shows some frightening looking white precipitate. When these mixtures were run in a gel (after vortexing so lots of the white particulate was loaded), all samples ran the same:
How much to load
Total Cell Protein
- use 5-8 or even up to 20 ug protein per well. (for Mini Protean with 10 well comb)
- Amanda runs 15 ug in skinny comb wells 2013/10/25
- 7.5 uL of turbid TB E. coli is about right for skinny wells. This becomes 10uL after adding 4X buffer.
Purified Protein
- ~40 ng seems about right. -JM
- This means the minimum concentration is ~5ng/uL. When you load 7.5uL in the skinny-comb gels (becomes 10uL after adding 4X buffer) you get ~40 ng in the gel.
- 4 to 8 uL*uM works well for ~50-70kDa enzymes. If the stock is 1uM, add 4-8uL. If it is 4uM, add 1-2uL. *JM 20 August 2014
In general
- To avoid edge effects, add 1x sample buffer to unused wells. (from protein gel electrophoresis tips and troubleshooting guide)
- [[User:Janet B. Matsen|JM] doesn't do this 2014/3
- If biomass is not limiting, prep a significant excess of protein, which will allow you to re-run if your gel turns out poorly for any reason.
- Maximum volumes:
- Load 12-15 uL, absolute max is 30 uL for 10-well comb in mini-protean
- Note: the amount you load affects the apparent size of the band:
- Load 12-15 uL, absolute max is 30 uL for 10-well comb in mini-protean
- If gel is overloaded or under loaded, run a new gel with a different amount of dyed & boiled lysate
How to prepare samples
- If protein normalizing:
- lyse cultures by sonication. Use 20 1 second pulses while tube is in ice, let the samples rest 5 minutes on ice, then sonicate again. Wipe sonicator stick between uses. Don't touch the tip while it is on.
- sonication allows you access to un-dyed lysed sample, unlike boiling with sample loading buffer as is described below.
- do Pierce BCA BSA assay to determine concentration. Run each sample with dilutions: 1, 1/10, 1/20, etc.
- Boil samples with sample buffer + beta-mercaptoethanol
- Optional: check efficacy of lysis with light microscopy (people usually don't do this.)
- Centrifuge all extracts extensively (20,000 x g for 15 min at 15°C) to remove any insoluble mater
- You can run gels with the soluble, insoluble, and/or crude (not centrifuged) lysates.
- lyse cultures by sonication. Use 20 1 second pulses while tube is in ice, let the samples rest 5 minutes on ice, then sonicate again. Wipe sonicator stick between uses. Don't touch the tip while it is on.
- If not normalizing:
- Boil cultures with concentrated sample buffer + beta-mercaptoethanol
- need to prepare this mix from concentrated loading dye (pre-made) and beta-mercaptoethanol. Make this mix fresh each day.
- If using Amanda's mix, mix 10 uL of this mix (proportions below) into every 25 uL of cell culture.
- If using BioRad's mix, "dilute the samples at least 1:2 with sample buffer", which I presume is 1 part cell culture + 2 parts mix.
- Heat your sample by either: (link)
- a) Boiling for 5-10 minutes (Works for most proteins)
- b) 65 degrees C for 10 minutes (If you have smearing using the above procedure)
- c) 37 degrees for 30 minutes (Membrane proteins or others that do not enter the gel otherwise may benefit from this type of sample preparation)
- consider centrifugation to remove insoluble??
- heat sample to 37oC to solubilize precipitated SDS??
- Boil cultures with concentrated sample buffer + beta-mercaptoethanol
Consolidated/Pictorial Protocol
Running the Gel
- Bio-Rad Mini-Cell Setup
- If only 1 gel, use buffer dam to replace second gel
- Position the gels with the shorter plate facing inward!
- Apply pressure on gel holder and gels as you close the tabs to seal the center compartment.
- Fill central compartment with running buffer
- should fill sample wells
- Pour more into the outer compartment to specified line
- Load gel
- Make sure you will be able to determine the orientation of your gel after it is stained. Asymmetry is good!
- Make sure to color/charge-match the cords to the power unit as the electrodes in the gel holder to the contacts in the lid.
- Run @ 60 V for ~15 min, then 200 V for ~ 20+ min.
- Note: ladder looks blurry while running through the stacking gel; don't be alarmed unless it still looks blurry in the resolving gel.
- Can skip the 60V step if you don't need a gorgeous gel
- Amanda runs 20 min at 200V, then checks frequently to make sure the protein doesn't run off the gel.
- Lower voltages can be considered if 200V leads to poor resolution.
- "[the] best typical conditions for electrophoresis are at 10-15 V/cm." (reference: protein gel electrophoresis tips and troubleshooting guide)
Storing Samples After Use
- Amanda was taught to flash-freeze (liquid N2) samples and store them at -80oC. Janet loves liquid N2 (particularly dumping it on the ground after use) so I follow along without question.
- Ladder is stored at -80oC. An aliquot of PageRuler Prestained that sat out at room temp for 1 week looked perfect in a gel, so don't worry about that ladder's stability.
Staining, Destaining, & Visualization
Old school bromphenol blue dye
- dye overnight or cycles of 1 min @ power 6 in the microwave
- microwave by Bo's bench. Let it vent a little in the hood between heating events.
- Return dye to container
- Rinse to remove residual dye
- Destain (2 - 4 cycles)
- The destain we use is just as potent when diluted 1:1 with tap water. I do this to save material and destain solution mixing time. -JM 12/2012
- Save the destain from the 3rd & 4th cycles, and re-use it for a future gel's 1st destain.
- Image with white light transillumination
- use the white light box, set the Gel Logic program to "white light transillumination", and use an exposure time of about 0.15 seconds.
- Using 1 second (and a tighter aperture) results in a blurry image
- Using less than ~ 0.15 seconds leads to inconsistent exposures. (The final image won't usually match the exposure level shown in the preview screen.)
- use the white light box, set the Gel Logic program to "white light transillumination", and use an exposure time of about 0.15 seconds.
New school GelCode Blue Safe Protein Stain
- Product Info:
- GelCode Blue Safe Protein Stain: A coomassie gel stain that's safer and less costly to ship.
- Pierce Item #1860957, Item# 24594
- How to use:
- Wash gel 3x in tap water (says Frances). Let it marinate ~10 min each time.
- Pour enough on to cover gel
- stain 30 min - overnight. (Frances likes overnight)
- Dump leftover stain down drain
- you might consider re-using bugger if your bands are fat
- Destain in water, but not overnight (too long in Amanda's experience)
Other Resources
- Modular recipe for PAGE pouring.
- You can download/copy the file and change the recipes as you like to get different gel percentages and batch sizes.
- Also includes a small scale (less than 2 mL) batch recipe on the 2nd sheet for troubleshooting failed polymerization or testing the functionality of the reagents.
- Background (Wikipedia)
- Instructions for PageRuler protein standard
- Instructions for BioRad Rainbow Std
- Mini-PROTEAN® Tetra Cell manual
- more comprehensive BioRad manual for SDS-PAGE (Janet's favorite)
- Criterion manual (for larger sized gels)
- video how-to (not vetted)
- article about polymerization from BioRad
Mistakes to Be Careful About
Prepping Gels
- using an "old" APS solution when making gels. The 10% weight/volume APS can be made fresh each time for best results. Don't use a solution that is more than a month or so old. The gels won't polymerize.
- not mixing the liquid gel mixture enough
- Janet pipettes up and down with a 10 or 15 mL pipette about 10 times, being careful not to bubble the solution.
- letting the gel dry too long after pouring the stacking gel (comb step)
- the very edges can shrivel up, which becomes a problem when you try to use those edge lanes
- taking the gel off the casting stand before it has polymerized
- entire sample will leak out
- If using commercially prepared gels, don't forget to remove the tape on the bottom of the gel! If you load your samples and turn on the power and see voltage but no current, check for tape removal (duh, Nicole)!
Preparing Samples
- using too much beta-mercaptoethanol in your sample buffer
- should have < 1% beta-mercaptoethanol in the mix after you add sample buffer to the
- too much reduction of cysteines is bad: will alter structure and even cleave proteins.
Running Gels
- sample sloshing out of the well you are trying to load and falling into a neighboring well
- Electrode buffer leaking out from between the two plates
- Gel will run very unusually or stop running. Keep an eye out!
- avoid by making sure the plates are well-seated
- Using Tris-HCl instead of Tris in the electrode buffer
- not having the lid to the running unit on all the way. --> poor electrical contact & blurry bands
- Using resolving gel Tris Buffer instead of the intended electrode buffer.
- The results of said mistake: JM 2013/01/04
Visualizing Gels
- Gels tear easily when warm.
- Be sure not to tear the gel as you take it out. It is good to wet the glass after you remove the cover glass plate.
All recipes except the staining & destaining solution are from the Mini-PROTEAN® Tetra Cell manual
10% (w/v) APS
- 0.10 g/1 mL diH2O = 100 mg/mL (prepare fresh daily for best results; most people do every few weeks.) (link)
Loading Buffer
- Mix pre-made concentrated loading buffer with fresh beta-mercaptoethanol prior to each use.
- TALON recipe: 5X SDS PAGE Sample Buffer
- 15% β-Mercaptoethanol (β-ME)
- 50% Glycerol
- 1.5% Bromophenol blue
- BioRad's loading buffer recipe:
- 3.55 mL deionized water, 1.25 mL 0.5 M Tris-HCl pH 6.8, 2.5 mL glycerol, 2.0 mL of 10% (w/v) SDS, 0.2 mL of 0.5% (w/v) Bromophenol Blue. Total volume = 9.5 mL.
- TALON recipe: 5X SDS PAGE Sample Buffer
- mix 50 uL beta-mercaptoethanol to 950 uL sample buffer prior to use.
- Scaled back 5x: 10 uL beta-mercaptoethanol + 190 uL 5x buffer
- scaled back 10x: 5 uL beta-mercaptoethanol + 95 uL buffer
- Dilute the sample "at least 1:2 with sample buffer" and heat at 95oC for 4 min to lyse the cells.
- Janet presumes this is 1 part sample per 2 parts buffer, but isn't sure (???)
- Amanda's recipe is a little different: 4x mix made of: 4 mL glycerol, 0.8 g SDS, 2.5 mL 1M Tris-HCl pH 6.8, 80 uL of bromophenol blue slurry (5 mg/mL = 0.5% (w/v)), H2O to 8 mL.
- Amanda mixes 160 uL 4X buffer, 8 uL beta-mercapto-ethanol, 32 uL H2O. Use 10 uL per 25 uL of cells. Recipe Comparison (to BioRad's)
- Can add up to 8M urea for really hydrophobic proteins
Running/Electrode Buffer:
- 10x SDS-PAGE (1 L) (BioRad catalog #161-0732) = 250 mM Tris, 1.92 M glycine, 1% SDS, pH 8.3
- Mix:
- Tris base 30.30 g DO NOT USE Tris-HCl
- Glycine 144.10 g
- SDS 10.00 g
- diH2O to 1 L
- Do not adjust the pH (~pH 8.3)
- Mix:
- Make 1L of 1x for use.
- Store at 4oC.
- This buffer is used while running proteins through the gel. Pour it in as the instructions for the box explain. Pour back into bottle for re-use afterward.
- It is best practice to not re-use electrode buffer. Our lab does, however, re-use the buffer ~ 4 (but sometimes up to 10) times.
Staining Buffer (Coomassie Brilliant Blue G-250):
- recipes vary. Usually there is about 0.5 g/L dye, and between 200 - 500 mL methanol per liter. 100 mL/L acetic acid is very common.
- 0.500 g Brilliant Blue, 500 mL methanol, 100 mL glacial acetic acid, 400 mL dH20. Mix well. Store at room temperature; can be reused 2-3 times.
- Amanda's recipe: 0.4 g of Coomassie Blue R 350 in 200 mL of 40% (v/v) methanol in water. Stir & filter (coffee filter is fine). Add 200 mL of 20% acetic acid in water (40 mL acetic acid in 160 mL water).
- note: Coomassie Brilliant Blue G-250 differs from Coomassie Brilliant Blue R-250 by the addition of two methyl groups. We use the G form. Read more about the R form here or at the bottom of this page.
Destaining Buffer:
- 30% methanol, 10% acetic acid, water
- some labs use much less methanol & acetic acid; some use plain water.
- Janet rinses in plain water before using our destaining buffer.
Acrylamide toxicity
- Acrylamide is toxic to your nervous system, and may be a carcinogen. The unpolymerized form is toxic, but the polymerized form is much less toxic. ALWAYS wear gloves and wipe up spills - once the solution dries, the dust can be inhaled. Interestingly, fried starchy/sugary foods naturally contain acrylamide, too.
- more than you want to know about acrylamide toxicity can be found here
The Lidstrom Lab PageRuler ladder best. (manual)
- The mobility of prestained proteins can vary in different SDS-PAGE buffer systems (manual)
- We load 4uL per well for 0.75mm thick gels; the manual suggests 5uL.
- You can't get use the ladders to approximate/quantify how much protein you added:
- "Thermo Scientific ladders are not designed for protein quantification. For quantification, we would recommend to use a protein of known concentration as a reference."
Other ladders:
Combs/Loading Volumes
- We currently only have the mini-protean gel running boxes. -JM 10/2012
- Hycult Biotech troubleshooting guide
- protein gel electrophoresis tips and troubleshooting guide includes troubleshooting after general tips
- nippongenetics guide
What is the best type of control to test expression of your plasmid?
- If you are using SDS-PAGE to test for expression of a plasmid, include an empty vector version of the strain you are looking for. This is good because you can look for your band, which hopefully doesn't overlap with a band that already exists in the host strain.
- This is also better than adding purified protein in a separate lane because the samples can run at different speeds. I'm not sure exactly why this happens, but here is an example:
How long are commercial PAGE gels good for after their expiration date?
We have used gels that expired 2-3 years before their use date. Specifically they were BioRad TGX gels. -JM 3/2015 They looked great!
How do you prepare soluble and insoluble fractions for SDS-PAGE?
Yakov Kipnis of the Baker lab wrote in 11/2014: "Soluble and insoluble fractions of cell lysate look distinct on gel (ratios of band intensities for various proteins is quite different), therefore if you see large portion of unlysed cells contaminating insoluble fraction distinction between soluble/insoluble becomes less pronounced. To make the comparison easier I typically run 3 lanes/sample (total cell lysate/soluble(sup after spin)/insoluble(pellet after spin) I always try to load similar amount of protein on a gel. For "total" and "soluble" it is straightforward, to help solubilize as much of "insoluble" as possible I resuspend pellet in small volume of 8-9 M urea (avoid GuHCl it destroys SDS gels) first and reconstitute to the same volume before taking small aliquot for gel sample. Example: total volume of the lysate 500 uL (take 10 uL for gel="total"), spin (take 10 uL="soluble"), remove supernatant as much as possible, add 20-50 uL 9M urea, resuspend pellet, dilute to 490 uL by water/buffer (take 10 uL="insoluble"). This way theoretically overlay of "soluble" + "insoluble" lanes should give you "total", if corresponding bands do not sum up there is a chance something unaccounted happened. Later you can skip "total" to save space on gel."
What do all of the reagents do? How long do they last?
- In loading buffer and often in gels (not necessary to include in gels; can be used in sample buffer)
- polymerize acrylamide
- Stability:
- Biorad customer service 10/2012: "TEMED breaks down through interaction with oxygen, which produces many products in small amounts. TEMED, if stored properly, excluding oxygen, should not break down. It can pool up under the liner of the cap, and since it has very low viscosity, can creep between the threads of the cap and those of the bottle. TEMED color is a good indicator of degradation. TEMED becomes a darker yellow color when degraded. Our APS is guaranteed for 1 year at room temperature in dry form (powder). APS should last indefinitely if stored desiccated. "
- brand new BioRad TEMED opened 11/2012 was clear, not yellow. The old bottle was clear-yellow.
- glycine
- carry charge in the opposite direction as the negatively-charged SDS-covered proteins
- methanol
- in wash buffer
- glycerol in loading buffer: helps sample sink
- Bromphenol Blue
All about the "stacking" layer of the gel
- The stacking layer originally was a lower density acrylamide with a lower pH. The goal of "stacking" was to gather all the proteins into a tight band so that they all start from the "starting line" at the same time, and appear more clearly after completion. The lower pH of the stacking gel is intended to affect how much the glycine in the running buffer is attracted to the proteins, which affects the proteins mobility. This video explains it if you can tolerate the corniness.
- There are, however, different ways to achieve stacking:
- lower density gel on top that isn't at a different pH
- Example: commercially available gels from BioRad. The fixed-percentage gels have only one buffer ingredient.
- The fact that having different pH values in the stacking and resolving gels isn't so important clarifies why home-made gels that initially have different pHs in the two layers still work fine weeks after they were poured when presumably the buffers converge to equilibrium.
- a gradient of acrylamide percentages
- A stacking gel is not necessary when using a gradient gel, as the gradient itself performs this function. (thermo scientific)
- lower density gel on top that isn't at a different pH
Is it important to degas my water + buffer + acrylamide mix before adding the APS and TEMED, as the manuals recommend?
- "We recommend that you degas the solutions to get rid of nitrogen. Sometimes, the nitrogen may come out of solution and form bubbles in your gels. " -BioRad customer support 10/26/2012 JM
- "Proper degassing and filtering of the casting solution is critical for both reproducibility of the polymerization (oxygen removal)" (link)
- This manual suggests the consequence is poor polymerization. If you don't experience poor polymerization, maybe you don't need to worry about degassing...?
- Also note this is not the justification customer support provided. (see above)
Why shouldn't we overlay the gels with butanol or isopropanol as they polymerize?
- "We do not recommend using butanol/isopropanol because these may degrade the glue on the spacer plates and the plastic of the casting frame." -BioRad customer support 10/26/2012 JM
Should I soak my gel or run them without samples before use?
- They say no: "I have not heard that soaking the gels in water will improve the run. Talking with my colleagues, we thought it would have been detrimental because the tris and chloride ions will diffuse out during this time and in theory, should have made your gel run poorly."
- We had an episode in October & November 2012 where the gels ran funny unless they were pre-run without samples, but this was resolved by replacing gel-pouring buffers someone made with commercial BioRad buffers. About the temporary issue:
- Ladder (and all other protein samples) ran fine until 1/2 way through the gel, after which they nearly halted and stacked on top of one another. Immediately after, a ~1 cm wide band that spanned across the gel was visible as it refracted light differently. This band disappeared after a while of soaking, leading me to believe it was a buildup of one of the buffer compounds. When such a gel was soaked overnight in water and ladder was loaded in an unused adjacent well, and it ran perfectly without bunching up. This was a very repeatable phenomenon with all gels used before the problem was solved.
- Potential problems with soaking or pre-running gels:
- You might ruin the stacking nature of the gel by altering the buffer within the acrylamide matrix. This essentially converts your gel to a continuous buffer gel system, known to give more blurry bands.
Is it ok to re-use my electrode buffer?
- Many people re-use it up to ~ 20 times, however, you should know that the manuals recommend single use and understand why. This may help.
- 2015 update: If you run at 200V+ for more than an hour or so, the buffer gets very viscous. We not longer use it when this happens.
- When we say "viscous" we mean that the bubbles that are formed in the liquid when adding DI water out of the spigot produces bubbles that don't surface quickly. The buffer also appears to have white filaments around this point of use.
How long can I store my acrylamide gels?
- "Tris-HCl resolving gels are prepared at pH 8.6–8.8. At this basic pH, polyacrylamide slowly hydrolyzes to polyacrylic acid, which can compromise separation. For this reason, Tris-HCl gels have a relatively short shelf life. In addition, the gel pH can rise to pH 9.5 during a run, causing proteins to undergo deamination and alkylation. This may diminish resolution and complicate post electrophoresis analysis." (reference)
- note: commercial Tris gels have a shelf life of 6 months to 1 year. Amanda & Ceci don't keep hand-poured gels for more than a few (~4) weeks. (JM 11/2012)
- JM uses gels that are up to 11 weeks old. They work perfectly. I haven't tested longer yet.
- It is possible the commercial gels have other means of extending shelf life.
- note: commercial Tris gels have a shelf life of 6 months to 1 year. Amanda & Ceci don't keep hand-poured gels for more than a few (~4) weeks. (JM 11/2012)
What is the difference between Coomassie R and Coomassie G?
- useful link
- note: Coomassie Brilliant Blue G-250 differs from Coomassie Brilliant Blue R-250 by the addition of two methyl groups. We use the G form. Read more about the R form here.
- R in R-250 stands for Reddish hue while G in G-250 for Greenish hue. R-250 is dark reddish blue/purple stain while G-250 gives lighter greenish blue stain.
BioRad once told Amanda:
- The G-250 form the colloidal particles in an aqueous solution. This is an advantage for staining a gel because the colloids tend not to stain the gel matrices, reducing the background problem. When the colloids come close to the proteins, the dye molecule is removed from the colloids by the nearby proteins due to the higher affinity of proteins to the dye.
- R-250, on the other hand, doesn't form the colloids. Rather, an individual dye molecule is dispersed in a solution. Therefore, the dye molecules can interact not only with proteins but with gel matrices freely, creating the background staining issue.
Should I throw my gel away if I forgot to add SDS to the resolving gel?
- Janet thinks no, for two reasons:
- The manual says "Bio-Rad's precast gels also do not contain SDS and can be used for native or denaturing PAGE." (citation)
- In 11/2012 I forgot to, and the proteins ran wonderfully, anyway.
Other Tips
Collecting samples in PCR strip tubes makes life nice!
- You can concentrate samples using the PCR strip spinner on Marina's bench. It must be balanced!
- You can do boiling-driven lysis in a thermocycler
- Samples don't take up a lot of room in the freezer.
- To write on them (for recording the date, and strip number if you have multiple) you can use a fine-tip sharpie and cover it with a ring of scotch-tape. Without tape, the writing tends to wear off.
- We are in love with individually removable caps that stay attached as is sold here.
- Samples don't splattering when you take of the caps, reducing cross-contamination and mess
- If you only have one cap open at a time when loading your gel, you can tell much more easily which sample you just loaded, reducing the possibility that you have a problem.
- Do, still, take each cap off slowly when opening them if there is liquid in the lid of the chamber. Often after shaking the strip to mix it, I open the cap quickly and splurt the blue sample on my clothes. It always rinses clean after adding water, but it is nicer if this doesn't happen.
Excess salt in SDS-PAGE samples causes fuzzy bands and narrowing of gel lanes toward the bottom of the gel
- If the ionic strength is very high, no bands will appear in the lower part of the gel (a vertical streak will appear instead) and the dye front will be wavy instead of straight. Deionize any sample with a total ionic strength over 50 mM using columns such as Micro Bio-Spin™ columns, which contain 10 mM Tris at a pH suitable for SDS-PAGE. (source)
Success or failure of any protein analysis depends on sample purity.
- Interfering substances that can negatively impact SDS-PAGE include salts, detergents, denaturants, or organic solvents (Evans et al. 2009). Highly viscous samples indicate high DNA and/or carbohydrate content, which may also interfere with PAGE separations. In addition, solutions at extreme pH values (for example, fractions from ion exchange chromatography) diminish the separation power of most electrophoresis techniques. Use one of the following methods as needed to remove these contaminants:
- Protein precipitation — the most versatile method to selectively separate proteins from other contaminants consists of protein precipitation by trichloroacetic acid (TCA)/acetone followed by resolubilization in electrophoresis sample buffer. A variety of commercial kits can simplify and standardize laboratory procedures for protein isolation from biological samples
- Buffer exchange — size exclusion chromatography is another effective method for removing salts, detergents, and other contaminants
Polyacrylamide is oxidative, and so disulfide bonds may be re-formed after proteins enter the gel
- also, oxidation of agents used to reduce any disulfide bonds in the original sample may introduce artifactual new components in the gel pattern. (source)
SDS can form micelles
- The SDS concentration is greater than the critical micelle concentration (cmc), so SDS is present as micelles. Micellar SDS bound to the tracking dye unstacks at high gel concentrations in disc electrophoresis; so a tracking dye that marks the moving boundary front at low gel concentrations fails to do so at high concentrations (source)
KCl causes SDS to precipitate
- If you samples contain KCl you should dilute them or methanol precipitate them and resuspend them in 1X sample buffer. With low concentrations of KCl (<200 mM) you can run them on the gel but you should load every lane with sample buffer containing the same concentration of KCl (even if they are blanks). This will help the gel run a little less anomalously. (source)
- BioRad sells most of the reagents we use. You can make most of them yourself, if you'd like.
- The "diaper" that goes under the gel casting stand:
- Janet called BioRad , UW EH&S, and VWR on 11/29/2012 to identify a special pad that is particularly good for absorbing liquid acrylamide mixtures used in pouring acrylamide gels. The idea is that such a pad would prevent the dried acrylamide from being re-released into the air and breathed by scientists. Unfortunately, this doesn't seem to exist.
- We ordered these


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Jan 09, 2017 | Carpet One Floor & Home
As a main focal point of the kitchen and bath, cabinets help to set the style of a room's overall design and lead the tone for other elements such as flooring, wall décor and window coverings. In 2017, cabinetry will continue to progress toward more thoughtful, higher-end designs, while offering better-than-ever functionality. With the help of industry experts and interior designs, we've compiled the elements that are expected to define cabinetry in kitchen and bath design and beyond.
Available in select stores, Carpet One Floor & Home offers an extensive kitchen and bath product line featuring quality cabinetry and countertops from well-known brands. Contact your local Carpet One Floor & Home for more information.

subset: commoncrawl


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From what it looks like, this will be the last season with nice and clean uniforms for the NBA..
LAS VEGAS — The NBA Board of Governors approved a new look for uniforms and a couple of changes in the use of video replay.
Commissioner David Stern met with the media Thursday after the Board of Governors held their annual summer meeting.
He said the league is looking to add possibly $100 million in revenue by permitting small advertising patches on uniforms starting in the 2013-14 season.
"The view is, that the teams would need a significant time; one, to sell the patch; and number two, for Adidas to manufacture the uniforms, because the patch that would be on the players' uniforms would also appear on the jerseys at retail," said Deputy Commissioner Adam Silver, who is handling the uniform change.
Silver said there are revenue projections from European soccer teams that the NBA could use.


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In this course in the Healthcare Marketplace specialization you will identify, define, and describe potential business and public policy solutions to the challenges facing society's growing demand for health services. Students will master a body of knowledge on the health care sectors major components through reading and reflection.
Healthcare has many different cultural components that will be discussed as historic trends as well as future demographic challenges. You will understand diverse philosophies and cultures within and across societies as they relate to healthcare. This outcome is particularly critical because of the tradeoffs needed to be assessed as medical technology advances faster than budgets and perhaps cultures are sometimes willing to tolerate.
At the end of the day, you will gain an appreciation for the role of creativity, innovation, discovery, and expression across disciplines in the development of new medical care solutions through and examination of the physician, medical technology and financing sub-sectors of the health economy.


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February 17, 1996 - August 16, 2009
I had to put my cat, Q-Tip to sleep today.
I remember adopting her when she was only 6 weeks old and so small.
She used to cry when she heard my car coming down the street and would greet me at the door.
I still wonder how someone less than 10 pounds managed to take over the entire bed. (And that I smooshed myself up against the wall less I disturb her beauty sleep.)
Her fur was soft as a feather, and she would tilt her head a bit to get what she wanted.
She liked to sit in the window and watch the birds, and plot her cute domination.
A little white tip of fur topped her tail...like a q-tip.
She woke up with me, sat on the LA Times while I read it, and would lick the remaining soy milk from my bowl of cereal.


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Rising consumer demand for local foods has changed the job description for ranchers like Doniga Markegard.
Markegard, co-owner of in San Gregorio, Calif., loves working with cattle, but she's not fond of the hours of phone calls and emails it can take to sell directly to a customer.
"What I want to be doing is the part I love — working with the animals and raising my kids on the ranch," says Markegard. "But I also need to be marketing our product, going to markets and talking with customers. There are a lot of administrative aspects to running a small family ranch, and they are time-consuming."
Now a San Francisco startup is looking to act as the middleman, handling the logistics of gathering and delivering local goods to consumers' doorsteps so small farmers like Markegard don't have to.
Good Eggs began a year ago as a place where local food producers could sell their foods directly to consumers online, says CEO Rob Spiro. But producers needed more.
"We kept hearing the same thing from the producers," Spiro tells The Salt. " 'This is great,' they told us, 'but as I become more successful, I'm becoming a full-time distributor.' "
So Spiro and his business partners decided to step in. "What we need is a last-mile delivery system for our producers," says Spiro.
The problem is that whether you live in San Francisco or Des Moines, Dallas or Wichita, the modern food system is based on economies of scale: To keep food inexpensive and delivered predictably, regardless of the season, you need mass production and the mass movement of goods from large-scale farm to national distributor to superstore.
But similar networks for moving locally produced foods to market are sorely lacking, according to a 2010 report from the USDA's Economic Research Service.
To that end, Good Eggs acquired three trucks and a warehouse and, as of last Thursday, it will now deliver fresh local fruits, vegetables, meats, seafood and prepared foods right to consumers' doors throughout the San Francisco Bay Area. It plans to create a similar food hub in Brooklyn this spring.
The Good Eggs system works like this: Consumers order from a wide variety of locally made, artisanal products online — from baby food to cheese, oranges to muffins. Items are then baked or harvested fresh to order and sent to the Good Eggs' warehouse, where each individual order is put together manually.
The idea is to keep costs down by using an Amazon warehouse model of efficient distribution — except nothing is stored there. The warehouse is used instead for aggregating goods on delivery days. By bringing all the products together in one location and distributing them together, Good Eggs hopes to relieve producers of the logistical headache of direct sales, while earning them higher profit margins than they get from grocery stores.
And while the price of jam on the Good Eggs site is far higher than a jar of Smuckers at Safeway, the cost of most goods is comparable to those at boutique markets like Whole Foods.
Yes, other companies have tried, with some success, to deliver groceries to homes and offices. Giant grocers like Safeway now offer home delivery. Fresh Direct makes home deliveries in the New York area, and Spud will drop off items to your front door in cities across the U.S. and Canada. But all of these companies function as full-scale grocers, and keep fully stocked warehouses full of nonlocal and nonfood items.
More similar is , with a mission to also deliver only local produce and meats directly to consumers in New York and California. But Farmigo only delivers to offices and other group drop-off points. Home delivery, the company found, was too costly to make it work.
Good Eggs is looking to get around that problem by charging $3.99 for home deliveries. Depending on whether a customer orders from the Good Eggs site or from a producer's own website, the company also takes a percentage cut from each sale.
An impressive team of investors and advisers are working with Good Eggs, including Alice Waters of Chez Panisse, Michael Dearing of Stanford and Damon Horowitz of Google.
Even so, the challenges of reconstructing a 21st century local food delivery system are formidable. How the company will deal with the logistical complexity of coordinating 120-plus producers and potentially hundreds of drop-off points, and how consumers will react to limited drop-off days and times, remains to be seen. (Some also question how far a food can travel while still being "local.")
"Creating new distribution channels between farmers and consumers is no easy feat," says Melanie Cheng of Berkeley, Calif., author of Building Regional Produce Supply Chains and founder of FarmsReach, a community-driven platform for improving business operations on the farm.
Picky shoppers, low profit margins and complex distribution logistics are inherent challenges in the system. "Good Eggs," she says, "have their work cut out for them."
Copyright 2020 NPR. To see more, visit https://www.npr.org.


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Though the St. Stephen's and St. Agnes Saints went nearly point for point with St. Albans early in their Tuesday matchup, the Bulldogs pulled away for a 65-46 win.
Matt Sniezek led the Bulldogs with 21 points, which was a season high for the sophomore guard. And St. Albans never scored less than 10 points a quarter en route to securing a 7-12 record with three matchups left in the regular season.
Senior Justin Goldsborough came up big again for the luckless 4-12 Saints, leading his teammates with 14 points. Sophomore Dominic Steward hit two field goals and a pair of 3-pointers to rack up 10 points in the loss.
The defeat comes on the heels of a Saints nonconference victory against Haverford on Saturday. In district play, the Saints have gone 0-7. They welcome backyard rival Episcopal on Friday before hitting the road to take on Bullis next week.


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The hoopla continues over Facebook's initial public offering of stock, with word that the social media giant has increased the size of Friday's IPO by 25 percent.
According to a statement filed with the Securities and Exchange Commission earlier today, the company now plans to sell 421 million shares — up from the previous estimate of 337 million. According to Reuters, at 421 million Facebook's IPO would be the third largest in U.S. history, after those of Visa Inc. and General Motors.
Tuesday, as we reported, Facebook said it was responding to high demand for the shares with an increase in the expected price, to between $34 and $38 per share from the original estimate of $28 to $35.
If the shares sell at the high end of the new range, Facebook would raise about $16 billion.
As we've also reported, though, it's going to be next to impossible for small investors to get their hands on the stock at its initial offering price — and there's word that General Motors is going to stop advertising on the website because it questions the effectiveness of such ads.
Related posts from our friends at Planet Money:
-- Pizza Delicious Bought An Ad On Facebook. How'd They Do?
-- Is Facebook Worth $100 Billion?
Copyright 2016 NPR. To see more, visit http://www.npr.org/.


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Off-White Laundy Slim Fit Pureed Pumkin T-Shirt
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Brands such as C.P. Company, Emporio Armani, Stone Island, Palm Angels, Valentino and many more all use Certilogo labels on their products, where using the code online you can check for authenticity. Moncler use their own system: code.moncler.com. All these labels can be found on the inside of the garment. Please note this is brand dependant.
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Oklahoma County's sheriff wants basketball fans and the Oklahoma City Thunder to know that streaking is illegal — but cheering, loudly, for the NBA team has yet to land anyone in the local jail.
If that doesn't sound like typical law enforcement advice, agency spokesman Mark Myers believes, that's a good thing.
Myers runs the sheriff's office Twitter account, which has become one of the most popular in the nation for mid-sized law enforcement agencies thanks to tweets ranging from cheering on the Thunder — "#Thunder down for WHAT?!!!!" — to taking followers along on DUI checkpoints and selfies of the sheriff.
"I've decided we want our brand to be more personable, and we want to have a conversation with people about what we're doing," said Myers, a former television reporter who is convincing the agency to look into new ways people absorb information.
"Since I came out of a news background, I have just been like, 'Wow, I can almost do a 24-hour news cycle about what the people in our agency are doing.'"
Law enforcement agencies around the country use social media to interact with the public. But Myers said the live-tweeting from Oklahoma County, home to Oklahoma City, has helped the agency develop an identity among residents who typically don't have much interaction with law enforcement — because they're not getting arrested.
Drinking and driving is no bueno. #CincoDeMayo
— Oklahoma Co. Sheriff (@OkCountySheriff) May 4, 2014
On Throwback Thursdays, a social-media trend where users post old photos, Meyer tweets century-old photographs of lawmen on horseback or his office's dive team in the 1980s. Fugitive Fridays host tweets about the area's most wanted criminals, while other messages are about severe weather or potential scams.
But non-police business dominate other tweets, 140-character messages employing hashtags and "at" symbols to help users find and follow each other.
"If you are running the @OKCMarathon tomorrow, you should be in bed...not watching the @okcthunder. Good luck Sunday!" Myers tweeted in April. And while his beloved Thunder were amid a losing streak, he tweeted, "Streaking is illegal in Ok County."
Such interactions have helped him attract and engage with the office's 21,000 official Twitter followers, making it the most followed mid-sized law enforcement agency behind only Maryland's Howard County police.
"If people are paying attention to you when you talk about the Thunder, think about how many people will be paying attention to you when there's actual really important information," Myers said.
Was that an earthquake or the Thunder shaking things up?
— Oklahoma Co. Sheriff (@OkCountySheriff) April 27, 2014
Myers began live-tweeting from drunken-driving checkpoints two years ago, and the number of followers jumped by 4,000 after the first night. Although he sometimes reveals the locations, Myers said he thinks people enjoy following along as people get busted.
"Tweet-alongs are replacing ride-alongs where people can see what we're doing," Myers said. "We're using that format for a few things, hanging out with our warrant squad like a modern-day posse, the DUI checkpoints and patrols."
Cindy Evans, a 48-year-old mother from Ratliff City, is among those followers of @OkCountySheriff.
"Sometimes their tweets are funny, but they're effective because lots of people are on Twitter and can help find the criminals when they might have trouble finding them otherwise," Evans said.
Sgt. Jennifer Wardlow, a spokeswoman at the Oklahoma City Police Department, said her agency likes what the sheriff's office is doing — but prefers to emphasize "business-related things" its social media feeds. Her agency has around 12,000 Twitter followers.


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debuted in the world of published books with what would become the Bigtime
books, a paranormal romance series set in fictional Bigtime, N.Y. The author
likens the series to "female-centered comic books without the art." The
Elemental Assassin series takes place in the Southern metropolis of Ashland, a
community rife with giants, dwarves, vampires, and elementals – including
assassin and restaurant owner Gin Blanco, with the power to control stone and ice,
and a love of sharp objects. Both of these series are for the over-eighteen
the young adult (YA) reader, Jennifer developed the Mythos Academy urban
fantasy series. The stories focus on seventeen-year-old gypsy Gwen Frost, who
has the gift of psychometry, and her education at the Mythos Academy, a school
for descendants of ancient warrior socities.
Frost", the fourth book in the Mythos Academy series is Jennifer's latest
release. The story features a potential relationship between Gwen and handsome
Logan Quinn. The fact Logan's father would just as soon see Gwen become a soon
forgotten memory of life might be a bit of a stumbling block. Landing in jail
on their first date for allegedly helping the sadistic Loki escape from
prison…. Sure beats dinner and a movie, but how do you plan a second date from
behind bars?
employs an easy-to-read style of writing in all of her work. Her characters are
well-drawn and interesting, and the plots original with just enough twist to
keep readers turning the page.
What was the inspiration behind the Mythos Academy series?
I've always enjoyed mythology and all the stories of the gods, goddesses,
warriors, and creatures battling each other and going on these epic quests. I
remember watching the old Clash of the Titans movie whenever we would have
movie days in school, and I read things like The Iliad and The Odyssey for
class assignments over the years.
day, I thought it would be cool to write my own mythology story with my own
characters and magic, put my own spin on things, and tell the story that I
wanted to tell. So that's what I did, and that was sort of the beginning of my
Mythos Academy series. So far, I'm having a blast writing the books.
The assassin Gin Blanco began as a killer for hire, but seems to be evolving
into more of a vigilante or seeker of vengeance. Why did you decide to change
her motivation for killing?
A) I've
always enjoyed reading about assassin characters in fantasy literature because
they can be everything from cold to calculating to crazy. It just seems like
there are so many different stories that you can tell with assassin characters.
Gin Blanco, I wanted to start out with a character who was cold, reserved, and
closed off, even from her friends and family. But as my Elemental Assassin
series has progressed, Gin has opened herself up more to her friends and
family, and that's one of the things that has changed her focus a bit. Gin is
still an assassin, but now she's more of an assassin-with-a-heart-of-gold who
helps folks who can't help themselves.
Fantasy stories require worldbuilding - the construction of the locale,
environment, demographics, society the characters interact in. You have stated
that you sometimes find authors spend too much time on worldbuilding and not
enough on the plot. What do you think the proper balance is?
A) Worldbuilding
is a big part of any fantasy book. I think one of the fun things about writing
fantasy books is coming up with a town, city, castle, or whatever and then
dreaming up all of the magic, creatures, and more that live in that place and
how they interact with each other.
plot is equally important. You can have a really creative magic world/system,
but without strong characters and a strong plot, it's really just an empty
town, city, or castle. It's important to strike a good balance between your
worldbuilding and the characters who drive your plots forward and hopefully
keep readers engaged in their adventures.
With three active series, are you planning to branch out into other stories in
other worlds? If so, what do you have in mind?
I have a couple of ideas for new fantasy books/characters/series that I would
like to explore. I think it would be cool to maybe write a YA series with more
of a fairy tale or epic fantasy feel. I'd also like to write in some different
genres and maybe try my hand at a spy thriller, a heist book, or maybe even a
western someday. I have more ideas than I could ever possibly have time to
Any parting thoughts for fans and potential new readers?
Folks can visit my website at www.jenniferestep.com
for more information on my books and to read excerpts. Folks can also follow me
on Twitter at @Jennifer_Estep or check out my author pages on Facebook and
for taking the time to interview me. I appreciate it. Happy reading, everyone!
DA Kentner is an
award-winning author www.kevad.net


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If you're looking to save some money on your holiday shopping purchases—or any online shopping for that matter—you should check out some of the web's best deal sites before you pull out the plastic and add that item to your cart. They can save you a ton of money at checkout, and many can even help you research the best buys before you even decide what you really want. We asked you last week which sites you thought were the best, and then took at look at the five best deal sites and put them to an all out vote. Now we're back to feature the winner.
With close to 50% of the overall vote, Slickdeals took the top spot by a wide margin. It's community powered deal listings, rapidly rotating and exclusive discounts, and wide popularity brought it to the top early in the voting and carried it through to the end.
In second place with over 16% of the overall vote was FatWallet, another largely community powered deal site that doesn't just cover online purchases and electronics, but also coupon codes and brick-and-mortar retailers. In third with close to 15% of the vote was Woot!, which many of you praised for its wide variety of daily deals on various items, but most importantly its community powered Deals.Woot! site, where bargains abound. In fourth with over 10% of the votes cast was Dealnews and its network of deal sites, regularly updating every hour of every day with fresh discounts, coupon codes, and bargains worth checking out. Finally, bringing up the rear with 9% of the vote was BradsDeals, a deal site that eschews the technology focus that the others have and offers up discounts, coupons, and bargains on apparel, jewelry, household items, and other goods both online and from name-brand retailers.
The Hive Five is based on reader nominations. As with most Hive Five posts, if your favorite was left out, it's not because we hate it—it's because it didn't get the nominations required in the call for contenders post to make the top five. We understand it's a bit of a popularity contest, but if you have a favorite, we want to hear about it. Have a suggestion for the Hive Five? Send us an email at [email protected]!


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Since we're discussing windows, let's talk (BrE) double-glazing--i.e. having two panes of glass in a window in order to reduce noise and loss of heat. The AmE equivalent would be double-paned windows, but the AmE term is much less commonly heard term than its BrE equivalent since double-glazed/paned windows don't offer the same level of home improvement in the US as in the UK. I've never lived in a house in the US (I've lost count of how many) that didn't have windows that are structured to have two layers, and I've never lived in a house in the UK (I've lived in four) that had more than one layer of glass in a very (BrE) draughty/(AmE) drafty frame. But this is not to say that most American houses have double-paned/glazed windows--far from it. Instead, they have a window frame with two tracks, with a window permanently in the inner track and the option of a mesh (AmE and AusE) screen or a storm window (i.e. of glass or similar material) for the outer track. In the part of the country I'm from, where it's very, very cold in winter and very, very hot in summer, a spring ritual is to (AmE or BrE) swap/(BrE) swop the storm windows for screens, with a predictable reversal of ritual in the fall/autumn.
Incidentally, when I'm asked what (besides people) I miss about the US, window screens are in the top three. One cannot enjoy a breezy warm evening in England without sharing it with every moth, wasp and mosquito in the neighbo(u)rhood. (The other two in the top three, since I expect you'll ask, are electrical outlets in bathrooms and butter wrappers that are marked with measurements, for easy baking. Electrical outlets are definitely number one--the other two change position with the seasons.)
And as long as we're discussing materials that you can look through, the plastic stuff is called perspex (or Perspex) in BrE and plexiglass (or Plexiglas--or some other combination of capitalization and esses) in AmE. Both of these are originally proprietary names--and another AmE proprietary name for this kind of stuff is Lucite. (Though I'd use Lucite when it's a thicker, less flexible piece--such as in a paperweight or the like.)


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Athenagoras I (Spyrou) of Constantinople
His All-Holiness Patriarch Athenagoras I (Greek: Αθηναγόρας Α' ), born Aristokles Spyrou (Αριστοκλής Σπύρου), was the 268th Successor to the Apostle Andrew and Ecumenical Patriarch of Constantinople from 1948 to 1972.
Early life
Patriarch Athenagoras was born in Vasilikón, near Ioánnina, Epirus, Greece, on March 25, 1886. The son of the village doctor, his mother died when he was only 13. He attended the Patriarchical Theological School of Halki, graduating in 1910. Upon graduating he was tonsured a monk and ordained to the diaconate, taking the name Athenagoras. He served as archdeacon of the Diocese of Pelagonia before becoming the secretary to Archbishop Meletius of Athens (and future Ecumenical Patriarch) in 1919. He was raised to the episcopate as the Metropolitan of Corfu in December 1922, while still a deacon.
Archbishop of America
In 1930, Metropolitan Damaskinos, after returning from a trip to the Greek Orthodox Archdiocese of America, recommended to Patriarch Photios II that Athenagoras be appointed Archbishop of North and South America. Damaskinos felt that Athenagoras was best suited to fix the many problems of the archdiocese, and Photios made the appointment on August 30, 1930.
When he assumed his new position on February 24, 1931, Athenagoras was faced with the task of bringing unity and harmony to a diocese that was racked with dissension between Royalists and Venizelists who had virtually divided the country into independent dioceses. To correct this he centralized the eccelesiastical administration in the Archdiocesean offices with all other bishops serving as auxiliaries, appointed to assist the archbishop, without dioceses and administrative rights of their own.
During his 18-year tenure, he actively worked with his communities to facilitate these reforms, in part by expanding the work of the clergy-laity congresses. He established the women's Philoptochos, the philanthropic arm of the Church, as well as an orphanage, St. Basil's Academy. He also founded the first Greek Orthodox seminary in America, the Holy Cross School of Theology. His capable leadership allowed him to withstand the early opposition he faced and eventually gain the love and devotion of his flock.
On November 1, 1948, Athenagoras was elected Patriarch of Constantinople, and was flown to Istanbul in the personal airplane of US President Harry Truman. As patriarch, he was actively involved with the World Council of Churches and improving relations with the Pope. His long reign ended with his death in Istanbul on July 7, 1972.
His 1964 meeting with the Pope Paul VI in Jerusalem led to the mutual lifting of the Bulls of Excommunication that resulted in the Great Schism of 1054. This was a significant step towards restoring communion between Rome and Constantinople. It produced the Catholic-Orthodox Joint Declaration of 1965, which was publicly read on December 7, 1965, simultaneously at a public meeting of the Second Vatican Council in Rome and at a special ceremony in Istanbul. The declaration did not end the schism, but showed a desire for greater reconciliation between the two churches. Nevertheless, not all Orthodox shared this sentiment, including Metropolitan Philaret, who wrote a response to the patriarch that same year.
Excerpts from Conversations with Patriarch Athenagoras:
- "I do not deny that there are differences between the Churches, but I say that we must change our way of approaching them. And the question of method is in the first place a psychological, or rather a spiritual problem. For centuries there have been conversations between theologians, and they have done nothing except to harden their positions."
- "Those who accuse me of sacrificing Orthodoxy to a b[l]ind obsession with love have a very poor conception of the truth. They make it into a system which they possess, which reassures them, when what it really is, is the living glorification of the living God, with all the risks involved in creative life. And we don't possess God; it is He who holds us and fills us with His presence in proportion to our humility and love. Only by love can we glorify the God of love, only by giving and sharing and sacrificing oneself can one glorify the God who, to save us, sacrificed himself and went to death, the death of the cross."
- "Orthodoxy, if it goes back to the sources of its great tradition, will be the humble and faithful witness to the undivided Church. The Orthodox Churches, in coming together themselves in mutual respect and love, will set a movement of brotherhood going throughout the Christian world, giving the example of a free communion of sister Churches, united by the same sacraments and the same faith. As to the Orthodox faith, centered as it is on liturgical praise and worship, and on holiness, it will bring the criterion of spiritual experience to ecumenical dialogue, a criterion which will allow us to disentangle partial truths from their limitations so that they may be reconciled in a higher plenitude of truth."
- "But we Orthodox: are we worthy of Orthodoxy? Up till the efforts we have made in recent years, what kind of example have our Churches given? We are united in faith and united in the chalice, but we have become strangers to one another, sometimes rivals."
Athenagoras I (Spyrou) of Constantinople | ||
Preceded by: ? |
Metropolitan of Corfu 1922-1931 |
Succeeded by: Alexander (Demoglou) |
Preceded by: Alexander (Demoglou) |
Archbishop of North and South America 1931-1948 |
Succeeded by: Michael (Konstantinides) |
Preceded by: Maximus V |
Patriarch of Constantinople 1948-1972 |
Succeeded by: Demetrios I |
External links
- St. Paul's Greek Church
- Catholic-Orthodox Joint declaration of 1965
- A Protest to Patriarch Athenagoras On the Lifting of the Anathemas of 1054 by Metr. Philaret of New York (December 2/15, 1965)
- Another Common Declaration of Pope Paul VI and the Ecumenical Patriarch Athenagoras I dated 28 October 1967
- Remembering Patriarch Athenagoras
- "Patriarch Athenagoras: Prophet of Love" by Bishop John (Kallos) of Thermon (Orthodox Research Institute)
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The exhibition "Reclamation: Gardens of Past Industry," featuring photography by faculty member Steve Nelson, will open on Friday, Feb. 22, and continue through Thursday, March 14, in the gallery of the De Pree Art Center at Hope College.
Nelson will deliver an artist talk on Friday, Feb. 22, at 4:30 p.m., followed by an opening reception from 5 p.m. to 6 p.m.
The public is invited to the exhibition, artist talk and reception. Admission is free.
The exhibition features photographs from Nelson's sabbatical travels to abandoned industrial sites in Michigan, exploring recurring themes of ruin and rebirth.
Nelson is an associate professor of art and chairperson of the department at Hope, where he has taught since 1989. His photographs have been exhibited widely, including in solo and group exhibitions in Chicago and New York.
He specializes in teaching photography at the college. In January 2008, he presented the seminar "Photography: Past and Present Tense" during the college's annual Winter Happening event. He was one of two faculty members with work featured in an exhibition in the De Pree gallery in 2007, and the gallery featured his work in a solo exhibition in 1997.
The De Pree Art Center is located at 160 E. 12th St., on Columbia Avenue at 12th Street. The regular gallery hours are Mondays through Saturdays from 10 a.m. until 5 p.m. and Sundays from 1 p.m. until 5 p.m. Gallery hours may be reduced during breaks and holidays. The gallery is handicapped accessible.


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perlbook - Books about and related to Perl
There are many books on Perl and Perl-related. A few of these are good, some are OK, but many aren't worth your money. There is a list of these books, some with extensive reviews, at http://books.perl.org/ . We list some of the books here, and while listing a book implies our endorsement, don't think that not including a book means anything.
Most of these books are available online through Safari Books Online ( http://safaribooksonline.com/ ).
The major reference book on Perl, written by the creator of Perl, is Programming Perl:
- by Tom Christiansen, brian d foy, Larry Wall with Jon Orwant
- ISBN 978-0-596-00492-7 [4th edition February 2012]
- ISBN 978-1-4493-9890-3 [ebook]
- http://oreilly.com/catalog/9780596004927
The Ram is a cookbook with hundreds of examples of using Perl to accomplish specific tasks:
- by Tom Christiansen and Nathan Torkington,
- with Foreword by Larry Wall
- ISBN 978-0-596-00313-5 [2nd Edition August 2003]
- http://oreilly.com/catalog/9780596003135/
If you want to learn the basics of Perl, you might start with the Llama book, which assumes that you already know a little about programming:
- by Randal L. Schwartz, Tom Phoenix, and brian d foy
- ISBN 978-1-4493-0358-7 [6th edition June 2011]
- http://oreilly.com/catalog/0636920018452
The tutorial started in the Llama continues in the Alpaca, which introduces the intermediate features of references, data structures, object-oriented programming, and modules:
- by Randal L. Schwartz and brian d foy, with Tom Phoenix
- foreword by Damian Conway
- ISBN 978-1-4493-9309-0 [2nd edition August 2012]
- http://oreilly.com/catalog/0636920012689/
You might want to keep these desktop references close by your keyboard:
- by Johan Vromans
- ISBN 978-1-4493-0370-9 [5th edition July 2011]
- ISBN 978-1-4493-0813-1 [ebook]
- http://oreilly.com/catalog/0636920018476/
- by Richard Foley
- ISBN 978-0-596-00503-0 [1st edition January 2004]
- http://oreilly.com/catalog/9780596005030/
- by Tony Stubblebine
- ISBN 978-0-596-51427-3 [July 2007]
- http://oreilly.com/catalog/9780596514273/
- by James Lee
- ISBN 1-59059-391-X [3rd edition April 2010]
- http://www.apress.com/9781430227939
- by Randal L. Schwartz, Tom Phoenix, and brian d foy
- ISBN 978-0-596-52010-6 [5th edition June 2008]
- http://oreilly.com/catalog/9780596520106
- by Randal L. Schwartz and brian d foy, with Tom Phoenix
- foreword by Damian Conway
- ISBN 0-596-10206-2 [1st edition March 2006]
- http://oreilly.com/catalog/9780596102067
- by brian d foy
- ISBN 978-0-596-10206-7 [1st edition July 2007]
- http://www.oreilly.com/catalog/9780596527242
- by Joseph N. Hall, Joshua A. McAdams, brian d foy
- ISBN 0-321-49694-9 [2nd edition 2010]
- http://www.effectiveperlprogramming.com/
- by Sam Tregar
- ISBN 1-59059-018-X [1st edition August 2002]
- http://www.apress.com/9781590590188
- by Tom Christiansen and Nathan Torkington
- with foreword by Larry Wall
- ISBN 1-56592-243-3 [2nd edition August 2003]
- http://oreilly.com/catalog/9780596003135
- by David N. Blank-Edelman
- ISBN 978-0-596-00639-6 [2nd edition May 2009]
- http://oreilly.com/catalog/9780596006396
- by Linchi Shea
- ISBN 1-59059-097-X [1st edition July 2003]
- http://www.apress.com/9781590590973
- by Jan Goyvaerts and Steven Levithan
- ISBN 978-0-596-52069-4 [May 2009]
- http://oreilly.com/catalog/9780596520694
- by Tim Bunce and Alligator Descartes
- ISBN 978-1-56592-699-8 [February 2000]
- http://oreilly.com/catalog/9781565926998
- by Damian Conway
- ISBN: 978-0-596-00173-5 [1st edition July 2005]
- http://oreilly.com/catalog/9780596001735
- by Mark-Jason Dominus
- ISBN: 1-55860-701-3 [1st edition March 2005]
- http://hop.perl.plover.com/
- by Jeffrey E. F. Friedl
- ISBN 978-0-596-52812-6 [3rd edition August 2006]
- http://oreilly.com/catalog/9780596528126
- by Lincoln Stein
- ISBN 0-201-61571-1 [1st edition 2001]
- http://www.pearsonhighered.com/educator/product/Network-Programming-with-Perl/9780201615715.page
- by Darren Chamberlain, Dave Cross, and Andy Wardley
- ISBN 978-0-596-00476-7 [December 2003]
- http://oreilly.com/catalog/9780596004767
- by Damian Conway
- with foreword by Randal L. Schwartz
- ISBN 1-884777-79-1 [1st edition August 1999]
- http://www.manning.com/conway/
- by Dave Cross
- ISBN 1-930110-00-6 [1st edition 2001]
- http://www.manning.com/cross
- by Steve Lidie and Nancy Walsh
- ISBN 978-1-56592-716-2 [1st edition January 2002]
- http://oreilly.com/catalog/9781565927162
- by Tim Jenness and Simon Cozens
- ISBN 1-930110-82-0 [1st edition August 2002]
- http://www.manning.com/jenness
- by Richard Foley with Andy Lester
- ISBN 1-59059-454-1 [1st edition July 2005]
- http://www.apress.com/9781590594544
Some of these books are available as free downloads.
Higher-Order Perl: http://hop.perl.plover.com/
You might notice several familiar Perl concepts in this collection of ACM columns from Jon Bentley. The similarity to the title of the major Perl book (which came later) is not completely accidental:
- by Jon Bentley
- ISBN 978-0-201-65788-3 [2 edition, October 1999]
- by Jon Bentley
- ISBN 0-201-11889-0 [January 1988]
Each version of Perl comes with the documentation that was current at the time of release. This poses a problem for content such as book lists. There are probably very nice books published after this list was included in your Perl release, and you can check the latest released version at http://perldoc.perl.org/perlbook.html .
Some of the books we've listed appear almost ancient in internet scale, but we've included those books because they still describe the current way of doing things. Not everything in Perl changes every day. Many of the beginner-level books, too, go over basic features and techniques that are still valid today. In general though, we try to limit this list to books published in the past five years.
If your Perl book isn't listed and you think it should be, let us know.


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Nugget Potatoes with Pesto |
The third Wednesday of every month has been designated Potato Ho-Down Wednesday. Two of my fellow potato Ho's Cathy of Noble Pig and Krysta over at Evil Chef Mom say it is so. Every Wednesday we will all be cooking up those scalloped potatoes, mashed potatoes, fries, curries, poutine......if potato is in the name we will be there!!! Being the Ho that I am this is my second entry this month.
1 kg (2 lbs) nugget potatoes, scrubbed, halved
125 mL (1/2 cup) each packed fresh basil leaves and ½ cup packed parsley
2 - 4 whole peeled garlic cloves
30 mL (2 tablespoons) pinenuts
60 mL (1/4 cup)sun-dried tomatoes drained and ¼ cup julienned
coarse black pepper to taste
3 mL (1/2 teaspoon) salt
60 mL (1/4 cup)extra-virgin olive oil
¼ cup grated Parmesan cheese (optional but droolworthy)
Drop the potato chunks into salted boiling water and cook until tender but not mushy. Drain.
Meanwhile, combine all remaining ingredients except Parmesan in food processor and process until well combined. Recipe makes about 10 tbsp (150 mL) of pesto. Use desired amount to toss with the potatoes in this recipe. Cover and refrigerate the remainder for up to 3 days for another use like Grilled Pizza.
Return the drained potatoes to the hot pan in which they were cooked and add desired amount of pesto. Toss until hot potatoes are well coated. Sprinkle with grated Parmesan if desired. Serve warm.
Serves 6
You are reading this post on More Than Burnt Toast at http://morethanburnttoast.blogspot.com. Excerpts and links may be used, provided that full and clear credit is given to the author/owner of More Than Burnt Toast. All rights reserved by Valerie Harrison.


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Now, the strike on Nairobi was noteworthy in part because of the group claiming responsibility. As David and Gregory mentioned, al-Shabab is a militant organization from nearby Somalia. Analyst Bronwyn Bruton of the Atlantic Council says a few years ago it would've had little reason to strike outside Somalia's borders. More recently, al-Shabab has been evolving, turned to new purposes by the influence of al-Qaida.
BRONWYN BRUTON: It emerged in 2005 in the wake of international efforts to create a government in Somalia.
INSKEEP: Which hasn't had a functioning national government in quite some time.
BRUTON: Yeah. Not since 1991, which basically was the year of a horrible famine and civil war, a time of intense violence. So al-Shabab mostly is a domestic insurgency, that is intending to remove this government, remove foreign influence from Somalia.
Unfortunately al-Qaida has used this domestic unrest as an opportunity to send operatives, international jihadists, who are essentially capitalizing on the suffering of the Somali people. And al-Shabab has been characterized as an affiliate, a proxy of al-Qaida, even though it's really a very small number of people within al-Shabab who are threats to the United States, who are interested in acting outside Somalia's borders
INSKEEP: So you have really local people with local concerns, and their fight has been globalized in the interest of Al-Qaida. That's what you see happening.
BRUTON: Yes, it's taken many, many years for that to happen. al-Shabab, as I mentioned, emerged in 2005. We're now in 2013.
INSKEEP: OK, before this attack on Kenya, on this shopping mall, what was al-Shabab's relationship to Kenya, people in Kenya, anything else?
BRUTON: Al-Shabab has had a symbiotic relationship with Kenya. Al-Shabab gets a lot of its funding and a lot of its political support from the Somali diaspora. And there's an enormous Somali diaspora in Nairobi, particularly in the community of Eastleigh. For that reason it's been conventional wisdom among analysts that al-Shabab would be very reluctant to attack Nairobi. Because if they did that, there would be a profound backlash against the Somali community there and support for al-Shabab crumble.
The fact that they have chosen to do this attack in Nairobi is actually an indication that there's been a decisive schism between these local actors and al-Shabab - which I've been talking about - and this group of foreign jihadists who've been at the very top of the leadership, but has had some bad luck in trying to actually pursued their jihadist's goals.
INSKEEP: So should we think of this shopping mall attack as the work of a faction of al-Shabab? Is that a good way to think of it?
BRUTON: Yeah. I mean al-Shabab has, on their Twitter feed, they've released a list of names of people actually perpetrated the attack. It hasn't been verified, but a lot of them are ethnically Somali people who've come from Sweden; one of them is from Syria. Only two of them actually list their country of origin as Somalia. These are hard-core jihadis and they're not necessarily interested in Somalia at all.
INSKEEP: Should we think of this attack as a way to assert control over the agenda and the image of al-Shabab? Could this almost be an internal political battle that is being played out in the shopping mall?
BRUTON: I think the likelihood that this transnational faction of Al-Shabab is interested in Somalia primarily, is very thin. On the other hand, Kenya is full of Western interests. And if al-Qaida wants to target America, which is obviously its reason for being, Kenya is the place to be. They would much rather be operating in Nairobi where they can hurt more people, and they can make more progress in their jihad than they could ever hope to accomplish in Mogadishu.
INSKEEP: Would you remind us, to the extent that it's known, how involved the United States has been in targeting and battling against al-Shabab?
BRUTON: Al-Shabab is low on a list of U.S. priorities, I think. If you look at the amount of money that the U.S. has been battling al-Shabab, it's less than $2 billion since 2007. But Kenya is the economic engine - it's the lifeblood of East Africa. There are diplomats there. There are aid workers there. There are tourists there. It is a vital hub for U.S. interests.
INSKEEP: Is there a danger here for the United States, because perhaps one of the reasons - if you're a terror group - that you would mount an attack like this in the shopping mall, is to suck in the United States in some way.
BRUTON: Absolutely, that was their goal in Somalia. Certainly it was al-Qaida's goal in Somalia to attempt to draw the U.S. into yet another quagmire, it didn't succeed. But they will certainly attempt to draw the U.S. into Nairobi. The United States will certainly feel pressure to assist in respond.
INSKEEP: Bronwyn Bruton is deputy director of the Atlantic Council's Africa Center.
INSKEEP: It's MORNING EDITION from NPR News. Transcript provided by NPR, Copyright NPR.


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It's customary to write an exit letter when leaving an organization after a long period of time, so here's mine. I have no desire to flame or insult anyone. You don't have to read this. It's for my own soul to find peace. I share it in the spirit of love, especially for those who have prayed for me as I have gone through this difficult transition period over the last few years. God bless you.
A Few Preliminaries
First and foremost, I want to thank the many individuals over the years who have taught me the gospel or who have allowed me to teach them. I cherish moments spent in gospel doctrine class, in Seminary, in the High Priest's Group and in Bishopric and High Council meetings when we have been edified together. There is nothing like being fed by the Lord through the scriptures.
Second, I am at peace with this decision. I confirmed it with the Lord in prayer. In fact, it was the Lord who continually urged this action upon me for the past three or four days. I know, I know. You may say, "Well, it wasn't the Lord urging you to resign your membership." I disagree. It was the honorable thing to do. It saved sixteen men many hours of time away from their families.
Third, I do not feel any different. I still feel the warmth and comfort of the spirit of the Lord. You may say, give it time, give it time. I hear you. I know there are things about membership in the LDS Church I will miss, but I will not miss the spirit of the Holy Ghost because he abides with me still, as does the priesthood authority I received from my father when I was but a lad.
A Few Questions Answered
Now for a couple of silly questions: Will I remove my garments? No. They still mean something to me. Will I attend the three-hour block each week? No. I will probably only attend Sacrament meeting with Carol. I will probably have my own sacrament meeting at home for a while until I can find a group with whom I can participate. Community is important. I seek a new community.
Will I continue to pay tithing? Yes. Carol and I discussed this. It is important to her to pay tithing to the LDS Church. I would prefer to pay tithing to a group where I know it is being used 100% to help the poor. But, and I know this sounds strange, unity with Carol is important to me. This is a difficult thing for Carol. Some women would divorce their husbands for leaving the Church.
I know Carol is hurt and disappointed. She is a fifth-generation Mormon. It means everything to her. She defends it with all the passion of a she bear. I have learned not to cross her. I am grateful she attended some of the lectures. I will make no attempt to entice her to follow me. I will only love her as best I can for a man who knows so little about the romantic needs of a good woman.
Don't Blame This on Denver Snuffer
To all those who now say, "See, I told you so. I warned you the moment you started reading and writing about those books from Denver Snuffer." Yes, you're right. You warned me. And you warned me and then warned me over and over again. Thank you for your loving concern. It did no good. I had a witness of the truth the moment I first read PtHG that night in January of 2012.
So am I now a follower of Denver Snuffer? No. I never was and never will be. I barely know the man. I know his writings and have been edified by them, but I am a disciple of Jesus Christ. Let no man come between you and your God. I suppose if there's one thing I do feel different already, it is the realization that I now get my marching orders – my daily priorities – only from the Lord.
Previous to this afternoon's letter of resignation, I was always concerned about, "What would my priesthood leaders think of this?" or "What does the LDS Church say I should be doing with my time?" Maybe that's my own fault or failing, but it's has been ingrained in me from a lifetime of hearing it drilled into my soul – follow the prophet. He knows the way. Thanks, but no thanks.
Follow the Prophet – Mantra Gone Too Far
I think I finally realized how bad it was one Sunday in High Priest's group when I answered a question posed by the instructor about marriage. I simply emphasized something I know we have been taught about temple marriages – that they are conditional upon being sealed by the Holy Spirit of Promise. This is a true doctrine. Always has been and always will be as far as I know.
You should have heard some of the comments I got from some of the brethren. And these were brethren I used to lead as the High Priest Group Leader. Somehow, it turned into a diatribe on the importance of following the prophet, that only he could interpret scripture or pronounce doctrine. It was as if they felt I was personally attacking them and their marriages. I was simply stunned.
It seems as if this has become the last stand for the LDS Church. It is a position of fear – follow the prophet. Please don't misunderstand. I am not attacking President Monson or whoever is the current president of the LDS Church whenever you read this. I simply thank God he has given me a mind of my own, that I can think for myself and I can ask God directly for guidance in life.
Study the Doctrine – But Not Too Much
I feel as if a great burden has been lifted from my shoulders and another one placed in its stead. I am OK with that. It is the burden of life. I have simply shifted it from being overly concerned with conformity to the orthodox LDS way of thinking and acting, to now being careful to think, believe, act and live in the manner intended by my Heavenly Father. I have shifted my priorities.
Was the LDS Church holding me back? Yes and no. It's the classic story. We are urged to study the gospel. When we do, we learn things that don't jive with what is currently being taught or practiced in the LDS Church. We are nothing like the church Joseph organized in 1830. There are too many traditions and beliefs taught in the correlated curriculum that are simply not right.
Agency – It Only Goes So Far
I can't tell you how many times I wrote about something I had studied and received a response, "You had better get your views in line with the Brethren or you're going to be in trouble." I got this over and over, from the least studied and newest member of the Church to Stake Presidents (not my current Stake President) and Bishops who read and comment on my blog. It's amazing.
Do I hold any animosity toward the church or its leaders? No. I thank them for all they have done for me, especially my local leaders who have loved me, prayed with me, counseled me and tried to get me to conform to the orthodox ways of modern Mormonism. I am certain I have offended some of my brethren in the High Priest Quorum stake-wide with my writings. They told me so.
I understand the concern expressed by associates on the High Council and Stake Presidency in my former stake. They are concerned for their children and grandchildren. I get that. I apologize if you feel I am to blame for some of them leaving the church. Don't put that at my feet. And for those in my current stake whose children have resigned – that was their decision and theirs alone.
Faulted for Believing Too Much
So what do I believe now? I believe in God. I know the Father lives and loves me. I know my Savior loves me and leads me. I have become more familiar with His voice over the past two years than at any time in my life. The Holy Ghost is real. He is the comforter. I appreciate that. I know God has a plan for my life that is not over. I went through hell last year to understand that.
I believe Joseph was a prophet of God. More than that, I know he was. The knowledge received of the Holy Ghost is burned into our souls. I have known Joseph was a prophet from my youth. My testimony of the Book of Mormon and other scriptures has not changed. We received the Book of Mormon the way Joseph said it came forth – translated by the gift and power of God.
Those Last Fingers of the Testimony Glove
The restoration was real. God wanted to prepare a people for the establishment of Zion. It did not work out in the early church. The higher priesthood was taken from the earth. The authority of the Aaronic remained. I received that authority from my father. I still have it. Resigning from the LDS Church does not take that away. Priesthood is not controlled by any earthly institution.
Obviously I no longer believe the LDS Church has what it claims. It saddens me the LDS Church seeks to enforce beliefs by disciplinary measures. Some of the policies in the Church Handbook are an abomination. They can destroy families. It also saddens me to learn of manipulation from the highest councils of the Church to control the affairs of local members and local leadership.
We have often been taught, and I have taught it myself, "While the people in the Church aren't perfect, the Church itself is perfect." I disagree. The LDS Church was divinely instituted but is not now what the Lord will use to establish Zion. Zion was never intended to be established by a large institution. Zion is to be established in small groups where there are no poor among them.
A Few Closing Thoughts
I often hear the complaint from LDS Church members, "They leave the Church but they won't leave it alone." I owe so much to the LDS Church. I judged my mother harshly when she walked away from the Church after twenty-five years. I was wrong. I have since asked for and received her forgiveness. She was my age or a little older when she left. I understand now what happened.
Mother loved the gospel. She was a disciple of Christ. She taught me to love the Savior and to love to study. She could not have been more devoted to the Church and a better example for me. She taught Gospel Doctrine and performed more temple ordinances than anyone I have known. Yet in the end, she could not find community within the narrow-mindedness of some members.
Following the traditions of the fathers is a real problem in the LDS Church. It keeps the members from progressing. It's good to safeguard some things that build family unity but not good to hold onto false beliefs and practices such as an over-reliance on "the Brethren" to tell us what's what. That's why my mother left after years of dedicated service. And now I have followed in her path.


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PHOWN summaries and recordsVirtual Museum View records in Virtual Museum formatCoverage map for all species Species totals, Nest stats per species and Observer totals Species summary , or General query Observer records, or View VM record Photo of the week, or PHOWN repeats, or PHOWN priorities |
PHOWN record summary
Enter new vm record to see a different record and hit 'Go'.
Species allocated by Coordinator: Record details entered by participant: (see all records here for this participant) Google map for this record (zoom in and switch to satellite view)
Interactive map: N4 pf -
Photos uploaded by observer
Large photos
783, Sociable Weaver Philetairus socius (see species summary here)Record status ACCEPTED Vm 9831 [bulk_socw_robert/_rene.xls::: 78931] Species Observer(s) Thomson RL Country, town, locus South Africa, Northern Cape, Hotazel,
2722ABLocality Tswalu Kalahari, col 210 Latitude, longitude -27.21368, 22.43339 [0 m accuracy] Date 2013/11/27 Notes Acacia erioloba, 1 falcon chambers, tree height 7, nest height 4, 1 nest part/s Nest count 18 Nest site tree
To see this map with all other PHOWN records, click here. Note that the map on this page will load very slowly and probably will only work if you use Chrome as a browser.
History of repeat colony counts
Note: repeats from the same day are not shown.vm Species code Date Nests Notes 5505 783 6/12/2012 13 Acacia eri, 2 nest parts; 1 collapsed part 9831 783 27/11/2013 18 Acacia erioloba, 1 falcon chambers, tree height 7, nest height 4, 1 nest part/s
Vm 5505
Vm 9831


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If you've ever heard your dentist or hygienist talk about "calculus," they're not referring to a higher branch of mathematics. The calculus on your teeth is something altogether different.
Calculus, also called tartar, is dental plaque that's become hardened or "calcified" on tooth surfaces. Plaque begins as soft food particles and bacteria that accumulate on the teeth, and more so if you don't properly clean your teeth every day. This built-up plaque becomes both home and food source for bacteria that can cause tooth decay or periodontal (gum) disease.
Because of this direct link between plaque and/or calculus and dental disease, we encourage everyone to perform two important oral hygiene tasks every day. The first is to floss between your teeth to remove plaque as you are unable to effectively reach those areas with a toothbrush. Once you loosen all the plaque, the other really important task is a thorough brushing of all of the tooth surfaces to remove any plaque that may have accumulated since the last brushing. Doing so every day will catch most of the softer plaque before it becomes calcified.
Once it forms, calculus is impossible to remove by brushing and flossing alone. That's why you should have regular cleanings performed by a dental professional. Dentists and hygienists have special tools called scalers that allow them to manually remove plaque and calculus, as well as ultrasonic equipment that can vibrate it loose to be flushed away with water.
In fact, you should undergo dental cleanings at least twice a year (or as often as your dentist recommends) even if you religiously brush and floss daily. Calculus forms so easily that it's nearly inevitable you'll accumulate some even if you have an effective hygiene regimen. Your dental team can remove hardened deposits of calculus that may have gotten past your own hygiene efforts.
If you haven't been consistently practicing this kind of daily hygiene, see your dentist to get a fresh start. Not only will they be able to check for any emerging problems, they can clean your teeth of any plaque and calculus buildup so that you'll be able to start with a "clean" slate.
Calculus can be tenacious, but it not impossible to remove. Don't let it set you up for an unhealthy experience with your teeth and gums.
If you would like more information on reducing plaque buildup, please contact us or schedule an appointment for a consultation.