以百合DNA为材料,对影响其RAPD反应体系中的6个主要因素(模板DNA、Mg^2+、TaqDNA聚合酶、引物、dNTPs、退火温度)进行优化,最终确定了百合RAPD反应的最佳体系(25μL)为:40ng模板DNA,1.5mmol/LMg^2+,1.0 U Taq DNA聚合酶,1.0μmol/L引物,200μmol/L dNTPs,引物E13的最适退火温度为37.3℃。该体系的建立为今后应用RAPD技术鉴定百合属植物的种质资源和遗传多样性研究奠定了基础。
The optimum RAPD reaction system for Lilium was established using RAPD amplification with six factors ( DNA template, Mg^2 + , TaqDNA polymerase, primer, dNTPs and annealing temperature). The optimum RAPD reaction system (25μL) contains 40 ng DNA template, 1.5 mmol/L Mg^2+ , 1.0 U TaqDNA polymerase, 1.0μmol/L RAPD primer and 200μmol/L dNTPs. The suitable annealing temperature of primer E 13 was 37.3 ℃ The research laid the foundation for RAPD analysis on studies of germplasm resources identification and genetic diversity in Lilium.