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云南红豆杉简并锚定微卫星-PCR反应体系优化研究

, PP. 739-743

Keywords: 云南红豆杉,简并锚定微卫星-PCR,反应体系,交互正交设计

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Abstract:

采用交互正交设计L64 (421 )对濒危植物云南红豆杉简并锚定微卫星-PCR反应体系分别进行了5因素( Taq酶;Mg2+ ; dNTP;DNA及引物) 4水平的优化筛选, SAS软件统计分析表明: 3个单因素( Taq酶;Mg2+ ; dNTP)和2个交互作用(Taq ×Mg2+ ,Mg2+ ×dNTP)对此反应体系有显著影响作用。云南红豆杉简并锚定微卫星- PCR反应的优化体系是:在20μL反应体系中含有1 ×PCR buffer, Taq酶4 U,Mg2+ ( 2. 0 mmol?L-1 ) , dNTP ( 0. 4 mmol?L-1 ) ,DNA 75 ng,引物(1. 0μmol?L-1 ) ,退火温度(54 ℃) 。交互正交设计既能快捷有效地筛选出最佳PCR反应体系,又能揭示试验因素及其交互作用对扩增的影响程度,分析结果更客观、准确,具有一定的应用价值。

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