黑木耳ISSR-PCR反应体系的正交优化

The Orthogonal Optimization of ISSR-PCR Amplification System in Auricularia auricula

采用正交试验设计的方法,对黑木耳ISSR-PCR(简单重复序列区间-多聚酶链反应)反应体系中的5种主要因素(Taq聚合酶,Mg2+,模板DNA,dNTP及引物)4个水平进行优化筛选,确立了适合黑木耳ISSR分析的优化反应体系(20μL),通过梯度PCR试验筛选得到相应引物的最佳退火温度。

The orthogonal design was used to optimize ISSR-PCR amplification system of Auricularia auricula in five factors （ Taq DNA polymerase, Mg^2 ＋ , DNA template, dNTP, and primer） at four levels respectively. A suitable ISSR-PCR system （20 uL） for A. auricula was established. The optimal annealing temperature of ISSR-PCR for A. auricula reaction was proposed by gradient PCR.