生物技术通报Issue(7)：113-116,4.
黄鳝ISSR-PCR反应体系的建立及条件优化
Optimization for ISSR Reaction System of Monopterus albus with Orthogonal Design
摘要
Abstract
Based on the genomic DNA of Monopterus albus, the factors influencing ISSR-PCR system were explored with the orthogonal design. The optimized ISSR-PCR system (25 μl) was determined as follows: 2.5 mmol / L Mg2+, 250 μmol / L dNTPs, 0.25 μmol / L primers, 1.0 U Taq DNA polymerase, 30 ng DNA template. The reaction program was set including initial denaturation for 5 minutes at 94℃, 40 cycles of denaturation for 40 seconds at 94℃,annealing for 40 seconds at 47 ～57℃, extension for 1.5 minutes at 72℃,with a final extension of 10 minutes at 72℃. The clear and reproducible amplification was obtained by the established ISSR reaction system.关键词
黄鳝/ISSR/正交设计/反应体系Key words
Monopterus albus ISSR Orthogonal design Reaction system分类
农业科技引用本文复制引用
杨浩,陈宏喜,陈欣,杨太有..黄鳝ISSR-PCR反应体系的建立及条件优化[J].生物技术通报,2009,(7):113-116,4.基金项目
国家大学生创新性实验计划项目(081047602),河南省教育厅自然科学研究项目(2006180017),河南省动物学重点学科资助,河南师范大学国家级生命科学实验教学示范中心资助 （081047602）
