摘要
以14个烟草品种为试材,初步建立了适合于烟草AFLP分析的银染分析体系:烟草基因组的DNA提取采用CTAB法;地酶切连接同步进行;预扩增选用E+A和M+C引物;选择性扩增选用E+3和M+3引物;扩增产物利用6%聚丙烯酰胺凝胶电泳,后用银染法检测。采用优化的体系,20对引物组合扩增出清晰的烟草AFLP图谱。
AFLP （Amplified Fragment Length Polymorphism） silver-staining analysis system was established with 14 tobacco cultivars. The results of several reactions in the analysis system were as following： tobacco genomic DNA was extracted by CTAB method; the double enzymes restriction and the adapter-ligation were completed at the same time; E＋A and M＋C primer combinations were used in pre-amplification while E＋3 and M＋3 primer combinations in selective amplification; the PCR reaction products were analyzed on 6% denatured polyacrylamide gel and then stained by silver. With the optimized system, the AFLP fingerprinting of 3 tobacco cultivars amplified by 20 primer combinations had been obtained.
出处
《山东农业科学》
2007年第5期38-41,44,共5页
Shandong Agricultural Sciences
关键词
烟草
AFLP分析
银染体系
Tobacco
AFLP
Silver-staining analysis system
