摘要
通过不同浓度Mg2+、dNTP、模板DNA、引物的组合试验、及不同浓度引物、TaqDNA聚合酶的单因素试验,筛选蜡梅最佳ISSR反应体系为:每20μL中含1×PCRbuffer、2.5mmol.L-1MgCl2、0.2mmol.L-1dNTP、0.3μmol.L-1引物、30ng模板DNA、1UTaqDNA聚合酶;同时通过梯度PCR试验确定了不同引物的适宜退火温度。
To optimize ISSR amplification system on Chimonanthus praecox, the effect of three factors,Mg^2＋ ,dNTP and DNA were studied on three levels respectively, the suitable concentration of primer and Taq DNA Polymerase were studied also. The results showed that optimum ISSR reaction system of Chimonanthus praecox was every 20μL reaction system containing 1 ×PCR buffer,2. 5mmol·L^-1MgCl2 ,0. 2mmol·L^-1dNTP, 0. 3μmol·L^-1 primer, 30 ng DNA template and 1U Taq DNA polymerase. The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR.
出处
《北方园艺》
CAS
北大核心
2007年第10期161-163,共3页
Northern Horticulture
基金
上海市科技兴农重点攻关资助项目[沪农科攻字(2005)第1-3号]
关键词
蜡梅
ISSR
反应体系
梯度PCR
Chimonanthus praecox
ISSR
Reaction system
Gradient PCR
