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猪基因组AFLP指纹图谱的构建
引用本文:任军,黄路生,Gary Eyens,高军,艾华水,陈克飞,丁能水,邓素华.猪基因组AFLP指纹图谱的构建[J].农业生物技术学报,2003,11(2):179-182.
作者姓名:任军  黄路生  Gary Eyens  高军  艾华水  陈克飞  丁能水  邓素华
作者单位:1. 江西农业大学江西省动物生物技术重点实验室,南昌,330045
2. Department of Pathology,University of Cambridge,United Kingdom
基金项目:国家自然科学基金项目(39860063)和国家"973"项目(G2000016102)资助.
摘    要:摘要:对猪(Sus scrofa)基因组AFLP指纹图谱构建的各项条件进行了优化,包括双酶切消化、接头连接、预扩增和选择性扩增、凝胶电泳银染技术及荧光标记检测法等,建立了稳定的猪基因组DNA的AFLP指纹图谱构建方法,采用E32/T32引物组合在45个猪种基因组混合DNA中检测到28个多态标记。

关 键 词:关键词:猪  基因组  AFLP指纹图谱
修稿时间:2002年3月18日

AFLP Markers for Pig Genomic DNA Fingerprinting
Gary Eyens.AFLP Markers for Pig Genomic DNA Fingerprinting[J].Journal of Agricultural Biotechnology,2003,11(2):179-182.
Authors:Gary Eyens
Abstract:Abstract: The optimization of AFLP technique for porcine(Sus scrofa)genomic DNA fingerprinting was studied, including enzyme digestion, adapter ligation, preamplification, selective amplification, denatured polyacrylamide gel electrophoresis, silver staining and multicolor fluorescent detection. The Eco RⅠ+AAC/TaqⅠ+AAC selective primer combinations labeled by IRD 800 and IRD 700 fluorescent dyes were used. Gel running was performed in both Licor 4200 DNA sequencer and 4.5% PAGE(2600 V, 45 W, 50 ℃), and the electrophoresis images were analysed by the AFLP QuantarTM Pro software of Keygene(the Netherland). The result showed that the template DNA concentrations did not affect the AFLP quality, templates with 250 ng, 500 ng and 1 000 ng DNA had the same AFLP fingerprinting. And 28 polymorphic markers were detected in the same pooled genomic DNA of 45 pig breeds (populations) by E32/T32 primer combinations.
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