{"title":"[利用动态磁珠富集大熊猫微卫星标记物]。","authors":"Fu-Jun Shen, Phill Watts, Zhi-He Zhang, An-Ju Zhang, Stephanie Sanderson, Steve J Kemp, Bi-Song Yue","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>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. coil JM109 competent cells. In total 260 positive clones were identified from 2 880 transformants in the libraries which were screened by gamma-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.</p>","PeriodicalId":23770,"journal":{"name":"Yi chuan xue bao = Acta genetica Sinica","volume":"32 5","pages":"457-62"},"PeriodicalIF":0.0000,"publicationDate":"2005-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Enrichment of giant panda microsatellite markers using dynal magnet beads].\",\"authors\":\"Fu-Jun Shen, Phill Watts, Zhi-He Zhang, An-Ju Zhang, Stephanie Sanderson, Steve J Kemp, Bi-Song Yue\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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. coil JM109 competent cells. In total 260 positive clones were identified from 2 880 transformants in the libraries which were screened by gamma-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.</p>\",\"PeriodicalId\":23770,\"journal\":{\"name\":\"Yi chuan xue bao = Acta genetica Sinica\",\"volume\":\"32 5\",\"pages\":\"457-62\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2005-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Yi chuan xue bao = Acta genetica Sinica\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Yi chuan xue bao = Acta genetica Sinica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
利用链霉菌素包被磁珠(Dynal)上的寡核苷酸探针进行杂交，获得了大熊猫含有STR序列的400 ~ 600 bp DNA片段。将富集的DNA连接到pGEM-T中，然后转化到E. coil JM109感受态细胞中。用γ - 32p标记探针对文库中2880个转化子进行筛选，共鉴定出260个阳性克隆。最终得到54条序列，并成功设计出37对大熊猫STR引物。结果表明，该方法是一种高效的微卫星标记分离方法。
[Enrichment of giant panda microsatellite markers using dynal magnet beads].
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. coil JM109 competent cells. In total 260 positive clones were identified from 2 880 transformants in the libraries which were screened by gamma-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.
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