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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (3): 288-292.

所属专题： 水产渔业

• 水产 渔业 • 上一篇    下一篇

鲤鱼AFLP-银染体系的建立与优化

李 慧萍,傅洪拓,李法君,乔 慧,姚建华   

  • 收稿日期:2008-09-11 修回日期:2008-12-22 出版日期:2009-02-05 发布日期:2009-02-05

Construction of AFLP Silver-staining Analysis System in Common Carp

  • Received:2008-09-11 Revised:2008-12-22 Online:2009-02-05 Published:2009-02-05

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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，本实验室从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。

关键词: 有机肥, 有机肥, 化学药剂, 小白菜, 产量, 品质

Abstract: AFLP(Amplified Fragment Length Polymorphism)analysis system was established with one breed of Commom Carp-Jian Carp. The results of AFLP silver-staining analysis system in Commom Carp was as follows: blood cell DNA Extraction, MseI and EcoRI double enzyme digestion at 37℃ for 5h; E+A and M+C as preamplification primers; 1：8 for E+3／M+3 selective primer in 25 PCR reaction volume, touch-down PCR procedure for the high efficiency. We got 8 polymorphic selective primers from 64 pairs, which will lay a foundation for future research in the development of Commom Carp breed resources.