南京林业大学学报（自然科学版）Issue(1)：15-20,6.DOI:10.3969/j.issn.1000-2006.2014.01.003
杉木SRAP-PCR体系优化
Optimization of Chinese fir SRAP-PCR system
摘要
Abstract
In order to optimize SRAP-PCR reaction system for Chinese fir,the orthogonal design L16(45) was considered with different concentrations of Mg2+, dNTPs, primer, Taq DNA polymerase and DNA template in SRAP-PCR reaction system for Chinese fir. The results of PCR were evaluated by visual and variance analysis,then the concentrations of Mg2+, dNTPs,primer were rectified with single-factor design. Finally,the results showed that the best reaction system was the combination in 20μL reaction solution,containing of Mg2+ 2.25 mmol/L,dNTPs 0.15 mmol/L,primer 0.4 μmol/L,Taq DNA polymerase1.5μmol/min and DNA template 60 ng,2μL10×PCR Buffer and ddH2 O to 20μL. The first optimal an-nealing temperature was 35℃,and the second step was 53 ℃. The optimized reaction system of Chinese fir described a-bove can get clear and stable bands.关键词
杉木/SRAP-PCR/体系优化Key words
Chinese fir/SRAP-PCR/system optimization分类
农业科技引用本文复制引用
王新民,郑仁华,陈金慧,赵亚琦,徐阳,王颖,施季森..杉木SRAP-PCR体系优化[J].南京林业大学学报（自然科学版）,2014,(1):15-20,6.基金项目
国家林业公益性行业科研专项项目（201004049）；国家自然科学基金重点项目（30930077）；江苏高校优势学科建设工程资助项目 （）
