摘要
利用正交实验设计,对芥菜ISSR-PCR反应的5因素4个水平进行研究,建立了芥菜ISSR-PCR反应的最佳体系,即在25μL反应体系中,模板DNA40 ng,引物10 pmol,10×反应缓冲液,dNTPs为0.5 m mol/L,TaqDNA聚合酶1 U,Mg2+2.0 m mol/L.并进一步进行梯度退火实验,找到了芥菜ISSR最适的退火温度为51℃.
The orthogonal design was used to optimize ISSR amplification system for mustard in five factors （Mg^2＋, Dntp, primer, Taq polymerase and DNA template） at four levels. An optimized ISSR reaction system was established, namely 25 μL reaction system containing 40 ng DNA template, 10 pmol primer, 10×PCR buffer, 0.5 mmol/L dNTPs, 1Utaq polymerase and 2.0 mmol/L Mg^2＋. The optimal annealing temperature for ISSR-PCR reaction was identified to be 51 ℃ by gradient PCR.
出处
《西南师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第1期115-118,共4页
Journal of Southwest China Normal University(Natural Science Edition)
基金
重庆市自然科学基金(2006BB1337
2006BB1334)
