摘要
利用正交试验设计,从Mg2+、dNTP、引物、甘油4种因素3个水平,对不结球白菜ISSR反应体系进行优化,确立了适合不结球白菜的ISSR反应体系:在20μL反应体系中,含1×PCRbuffer,1.5mmol·L-1MgCl2,200μmol·L-1dNTP,0.5μmol·L-1引物,1%甘油,30ng模板DNA,1UTaqDNA聚合酶.通过梯度PCR测验,确定了适宜的退火温度.
In this study, orthogonal design was used to optimize ISSR amplification system on Brassica campestris ssp. chinensis in four factors (Mg^(2+), dNTP, primers, glycerine) at three levels respectively. A suitable ISSR reaction system was established, namely 20 μL reaction system containing 1×PCR buffer, 1.5 mmol·L^(-1) MgCl_2, 200 μmol·L^(-1) dNTP, 0.5 μmol·L^(-1) primer, 1% glycerine, 30 ng DNA template and 1U TaqDNA polymerase. The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR.
出处
《西北植物学报》
CAS
CSCD
2004年第5期899-902,共4页
Acta Botanica Boreali-Occidentalia Sinica
关键词
不结球白菜
ISSR
正交设计
梯度PCR
Brassica campestris ssp.chinensis
ISSR
orthogonal design
gradient PCR
