| 鲤鱼AFLP-银染体系的建立与优化 |
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| 引用本文： | 李 慧萍,傅洪拓,李法君,乔 慧,姚建华.鲤鱼AFLP-银染体系的建立与优化[J].中国农学通报,2009,25(3):288-292. |
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| 作者姓名： | 李 慧萍 傅洪拓 李法君 乔 慧 姚建华 |
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| 作者单位： | 1. 南京农业大学无锡渔业学院,江苏无锡,214081
2. 中国水产科学研究院淡水渔业研究中心,农业部水生动物遗传育种和养殖生物学重点开放实验室,江苏无锡,214081 |
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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。
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| 关 键 词： | 有机肥 有机肥 化学药剂 小白菜 产量 品质 |
| 收稿时间： | 2008-09-11 |
| 修稿时间： | 2008-12-22 |
Construction of AFLP Silver-staining Analysis System in Common Carp |
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| Li Huiping,Fu Hongtuo,Li Fajun,Qiao Hui,Yao Jianhua.Construction of AFLP Silver-staining Analysis System in Common Carp[J].Chinese Agricultural Science Bulletin,2009,25(3):288-292. |
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| Authors: | Li Huiping Fu Hongtuo Li Fajun Qiao Hui Yao Jianhua |
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| Institution: | Li Huiping, Fu Hongtuo, Li Fajun, Qiao Hui, Yao Jianhua (1.Wuxi fisheries College, Nanjing Agricultural University, Wuxi Jiangsu 214081; 2.Key Open Laboratory for Genetic Breeding of Aquatic Animal and Aquaculture Biology, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi Jiangsu 214081) |
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| 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. |
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| Keywords: | AFLP |
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