红花檵木RSAP-PCR反应体系的建立与优化

Establishment and optimization of RSAP-PCR reaction for Loropetalum chinese var. rubrum

分别采用单因子试验和正交设计试验对影响红花檵木RSAP-PCR反应体系的5个因素(DNA模板量、Mg2+浓度、dNTPs浓度、引物浓度、Taq酶量)在4个水平上进行优化.结果表明,2种方法得到的影响因素的最优水平存在差异,通过综合比较与分析,选取红花檵木RSAP-PCR最优反应体系为:在25μL的反应体系中,模板量70ng,Mg2+浓度1.50mmol/L,dNTPs浓度0.25mmol/L,引物浓度0.20mmol/L,Taq酶量2.50U.

Both a single-factor test and orthogonal-design test were conducted to optimize RSAP-PCR amplification system on Loropetalum chinese var. rubrum, at four levels of five factors （concentration of DNA, Mg^2＋, dNTPs, and primer, and contents of Taq DNA polymerase） respectively. The results showed that there were differences between the two methods at suitable levels of factors, and a suitable RSAP-PCR reaction system was established, namely, 25 μL reaction system containing 70 ng DNA template, 1.50 mmol/L Mg^2＋, 0.25 mmol/L dNTPs, 0.20 mmol/L primer, 2.50 U Taq DNA polymerase, through comprehensive comparison and analysis.