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[DNA fragment isolation of genomic microsatellite in Pinellia ternata by magnesphere].

OBJECTIVE
To develop microsatellite primers with the method of isolation of genomic microsatellite in Pinellia ternata by magnesphere.


METHOD
By taking advantage of the high binding affinity of biotin to streptavidin, microsatellite probe of the 5' end biotin was combined with magnesphere paramagnetic particles, and then combinations were hybridized with the digested P. ternata DNA fragments in which both ends of them connected with special adaptor, the other DNA fragments were eluted out. The microsatellite library was established. The crossed fragments were used as the template to conduct PCR amplification with the adapter sequences as primers, the products were cloned directly, subsequently screened by bacterium liquid PCR, and DNA sequencing was carried out.


RESULT
Fifteen microsatellites of P. ternata were obtained, development efficiency was 93.75%.


CONCLUSION
The result demonstrates that magnesphere is a fast and efficient method to develop microsatellite.