论文部分内容阅读
利用正交设计L_(16)(4~5),对酥瓜ISSR-PCR反应体系的5个影响因素(引物,d NTPs,Taq DNA聚合酶,Mg2+和模板DNA)在4个水平上进行优化试验,并在36℃~56℃范围内摸索退火温度,建立适合酥瓜ISSR-PCR反应体系,结果表明,在20μL反应体系中,含有引物0.2μmol/L、d NTPs 0.3 mmol/L、Taq DNA聚合酶1.2 U、Mg2+1.0 mmol/L、DNA模板70 ng、10×Buffer 2.0μL为最佳反应体系,ISSR-PCR扩增程序中最佳退火温度为52.5℃。该体系为酥瓜种质资源的遗传多样性分析评价提供了帮助。
The orthogonal design of L_ (16) (4 ~ 5) was used to optimize five factors (primers, dNTPs, Taq DNA polymerase, Mg2 + and template DNA) in the ISSR- The results showed that 0.2 μ mol / L of primers, 0.3 mmol / L of d NTPs and Taq DNA were prepared in a 20 μL reaction system. Polymerase 1.2 U, Mg2 + 1.0 mmol / L, DNA template 70 ng, 10 × Buffer 2.0 μL for the best reaction system, ISSR-PCR amplification program, the best annealing temperature was 52.5 ℃. The system provided a help for the analysis and evaluation of the genetic diversity of Canken germplasm resources.