药用半枫荷基因组总DNA提取及ISSR-PCR反应体系优化

Total DNA Extraction and ISSR-PCR Reaction System Optimization of Semiliquidambar cathayensis

利用正交试验设计方法,对影响ISSR-PCR反应的Mg2+、dNTPs、引物和Taq DNA聚合酶等因素进行优化的结果表明:在20μL PCR反应体系中含10×PCR Buffer,0.018μmol/L引物,4.0 mmol/L Mg2+,15 ng/μL模板DNA,1.5 U Taq DNA聚合酶,4种dNTPs各0.15 mmol/L为半枫荷ISSR-PCR最适反应体系。

In order to establish the optimal system of ISSP-PCR reaction for Semiliquidambar cathayensis,the parameters including template DNA,primers,dNTPs,Mg2＋ concentration,dose of Taq DNA polymerase were systematically analyzed.The results showed that the optimal ISSR-PCR reaction was carry out in a volume 20 μL containing 10×PCR Buffer,0.018umol/L Primer,4.0mmol/L Mg2＋,1.5U Taq DNA polymerase,4 dNTPs each 0.15 mmol / L.