正交设计优化柿属植物SSR-PCR反应体系  

Optimization for SSR-PCR reaction system of Diospyros L.trees by orthogonal design method

作　　者：梁玉琴[1] 李芳东[1] 傅建敏[1] 乌云塔娜[2] 吴硕[1] 

机构地区：[1]中国林业科学研究院经济林研究开发中心,河南郑州450003 [2]中南林业科技大学经济林育种与栽培国家林业局重点实验室,湖南长沙410004

出　　处：《经济林研究》2011年第4期17-22,共6页Non-wood Forest Research

基　　金：林业公益性行业科研专项项目"柿属植物种质资源保护与选育技术研究"(200904041)

摘　　要：为了快速有效地确定柿属植物SSR-PCR反应体系,给柿属植物遗传多样性的SSR标记研究奠定基础,采用正交设计的方法,利用2×Taq PCR Master Mix,将Taq酶、dNTPs和Mg2+3个因素作为整体,综合评价其对柿属植物SSR-PCR反应体系的影响。结果显示:反应各因素水平对PCR反应均具有显著影响,其中Mix影响最大,引物次之,模板DNA最小;确定柿属植物SSR-PCR反应体系为模板DNA 2μL(40～100 ng)、2×TaqPCR Master Mix 10μL、引物1μL,总体积25μL;反应体系具有较高的重复性和稳定性,筛选出20对多态性较高的柿属植物SSR引物。In order to rapidly and effectively determine SSR-PCR reaction system of Diospyros L.trees and lay a foundation for the studies on genetic diversity of Diospyros L.,the effects of Taq DNA polymerase,dNTPs and Mg2＋ as a whole on SSR-PCR reaction system of Diospyros L.trees were evalued synthetically,using 2×Taq PCR Master Mix by orthogonal design method.The results showed that the effects of every levels of each factor on PCR reaction were all significant.The most remarkable factor was 2×Taq PCR Master Mix,the least one was DNA template and the middle one was primer.The best SSR-PCR reaction system for Diospyros L.trees was DNA template 2 μL（40-100 ng）,2×Taq PCR Master Mix 10 μL,primer 1 μL with the totoal volume of 25 μL.The PCR reaction system had high repeatability and stability,and 20 pairs of SSR primers with high polymorphism were gotten.

关 键 词：柿属植物 正交设计 SSR-PCR反应体系 2×Taq PCR MASTER MIX 

分 类 号：S665.2[农业科学—果树学] S718.46[农业科学—园艺学]