摘要
以鲤科鱼类建鲤为材料,对扩增片断长度多态性(AFLP)反应体系进行优化,建立适合于鲤鱼的AFLP反应体系。通过对AFLP常规流程中基因组DNA提取、酶切-连接、预扩、选扩及电泳-银染过程的优化,初步建立了适合鲤鱼的AFLP分析体系:采用鲤血细胞DNA提取;MseI和EcoRI37℃双酶切5h;预扩引物为E+A和M+C;选扩引物E+3与M+3浓度配比为1:8,为提高引物与模板的结合效率,选扩程序均采用touch-down程序;最后结合经典AgNO3-Na2CO3银染显色。通过优化的AFLP程序,从64对引物中筛选出8对多态性引物,为进一步的鲤鱼种质资源研究奠定基础。
AFLP （Amplified Fragment Length Polymorphism） analysis system was established with one breed of Common Carp - Jian Carp. The results of AFLP silver-staining analysis system in Common Carp was as follows： blood cell DNA Extraction, MseI and EcoRI double enzyme digestion at 37 ℃ for 5 h; E＋A and M＋C as pre-amplification primers; 1 ： 8 for E＋3/M＋3 selective primer in 25 PCR reaction volume, touch-down PCR procedure for the high efficiency. Eight polymorphic primers were screened out of 64 pairs, which will lay a foundation for future research in the development of Common Carp breed resources.
出处
《中国农学通报》
CSCD
北大核心
2009年第3期288-292,共5页
Chinese Agricultural Science Bulletin
基金
国家"十五"科技攻关项目"全雌建鲤选育技术研究"(2004BA526B0109)
关键词
鲤鱼
AFLP
银染体系
common carp, AFLP, silver-staining system
