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桃SRAP-PCR反应体系的建立与优化

DOI: 10.7668/hbnxb.2008.S2.046, PP. 201-204

Keywords: ,SRAP,优化,正交试验

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Abstract:

建立适宜桃基因组DNA的SRAP-PCR扩增体系,为桃基因图谱的构建和分子标记打下基础。以桃基因组DNA为模板,通过正交试验设计,从dNTPs、Mg2+、Taq酶、引物、模板5种因素4个水平对桃SRAP-PCR反应体系进行优化,所建立的体系为25μL:dNTPs为0.12mmol/L,Mg2+为4mmol/L,Taq酶2U,引物为0.3mmol/L,模板DNA50ng。PCR反应程序为:94℃预变性5min;94℃变性lmin,35℃复性lmin,72℃延伸lmin,5个循环;94℃变性lmin,50℃复性lmin,72℃延伸lmin,35个循环,72℃延伸10min。

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