Dynal磁珠富集大熊猫微卫星标记

Enrichment of Giant Panda Microsatellite Markers Using Dynal Magnet Beads

应用Dynal磁珠生物素标记的微卫星探针与大熊猫基因组酶切片段杂交,捕获400～600bp含有微卫星序列的DNA片段,连接到pGEM T载体中,构建富集微卫星序列的小片段插入文库。应用γ32P标记的探针筛选文库,从2880个转化子中获得了260个阳性克隆。对54个序列进行了测序,并成功地设计了大熊猫微卫星引物37对。该方法能有效提高筛选微卫星标记的效率。

The 400~600 bp DNA fractions of giant panda containing STR sequences were captured by hybridization with the oligonucleotide probes attached to streptavadin coated magnetic beads (Dynal) .The enriched DNA were ligated into pGEM-T and then transformed into E.coli JM109 competent cells.In total 260 positive clones were identified from 2 880 transformants in the libraries which were screened by γ-^(32)P radiolabelled probes.Finally,we got 54 sequences and successfully designed 37 pairs of STR primers for giant panda.The results showed that this method is very efficient to isolate microsatellite markers.