摘要
利用正交设计L16(45)对异色瓢虫(Harmonia axyridis Pallas)ISSR-PCR反应体系的5个因素(Taq酶、Mg2+、模板DNA、dNTPs、引物)在4个水平上进行优化试验,试验结果用DPS软件进行分析,建立了异色瓢虫ISSR-PCR反应的最佳体系,即在20μL体系中模板150ng、引物0.5μmol/L、dNTPs0.15mmol/L、TaqDNA聚合酶1.5U、Mg2+1.25mmol/L。并对反应体系进行梯度退火试验,得到最佳退火温度为53.3℃。
Five factors of ISSR-PCR amplification system, Taq DNA polymerase, Mg^2＋, DNA template, dNTPs and primer, were optimized at four levels by the orthogonal design using Harmonia axyridis （Pallas） as the experimental material. Results of PCR were analyzed by DPS software. The optimal ISSR-PCR system for H. axyridis was obtained as DNA template 150 ng, primer 0.5μmol/L, dNTPs 0.15 mmol/L, Taq DNA polymerase 1.5 U, and Mg^2＋ 1.25 mmol/L in a 20 μL reaction system. The suitable annealing temperature for the ISSR-PCR reaction was 53.3 degrees C according to gradient PCR.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2008年第12期46-48,共3页
Journal of Northeast Forestry University
关键词
异色瓢虫
ISSR
正交设计
分子标记
天敌昆虫
Harmonia axyridis
ISSR
Orthogonal design
Molecular marker
Natural enemies
