红麻SRAP-PCR反应体系优化及多态性引物的筛选  

Optimization of SRAP-PCR Reaction System by Orthogonal Design and Screening of Polymorphic Primers for Hibiscus cannabinus L.

作　　者：武耀龙[1] 李德芳[1] 黄思齐[1] 李建军[1] 唐慧娟[1] 陈艳翠[1] 陈安国[1] 

机构地区：[1]中国农业科学院麻类研究所,湖南长沙410205

出　　处：《中国麻业科学》2014年第1期1-7,共7页Plant Fiber Sciences in China

基　　金：国家麻类产业技术体系

摘　　要：采用正交试验L16(43)设计,以dNTP、引物和Taq DNA聚合酶3种因素4个水平,并设置了8个模板DNA浓度梯度。扩增效果表明,红麻的SRAP-PCR最佳反应体系为2μL10×Taq Reaction Buffer、20ng模板DNA、dNTP 220μmol/L、引物0.35μmol/L、Taq DNA聚合酶0.5U,总体积为20μL。同时对256对SRAP引物进行扩增,筛选出条带清晰、多态性较好的引物100对。该反应体系及100对多态性引物组合应用于今后红麻的遗传多样性、品种鉴定、亲缘关系、遗传图谱构建、QTL定位等研究。An orthogonal experiment L 16 （ 43 ） including three factors （ dNTP, primer and Taq DNA Polymerase ） and four levels was designed to optimize SRAP -PCR reaction system of Kenaf.Be-sides, we set up 8 concentration gradients of DNA template to test its influence on amplifying results.At last, we optimized the SRAP-PCR reaction system for Kenaf.It contained 2μL 10 ＆#215;Taq reaction buff-er, 20ng DNA template, 220μmol/L dNTP concentration, 0.35μmol/L primer concentration, 0.5U Taq DNA Polymerase , 20μL of the cumulative volume.In addition, 100 primer combinations were se-lected from 256 SRAP primer combinations due to their distinct Gershgorim bands and high polymor -phism.The optimal SRAP reaction system and 100 primer combinations would contribute to the basis for evaluating genetic diversity , identifing varieties , analyzing genetic relationship , building linkage map and QTL map for Hibiscus cannabinus L.

关 键 词：正交试验设计 红麻 体系优化 引物筛选 

分 类 号：S563.5[农业科学—作物学]