银胶菊的SRAP-PCR反应体系优化及引物筛选  

Optimization of SRAP-PCR Reaction System in Parthenium hysterophorus and Primers Screening

作　　者：高翔[1] 关亚丽[1] 

机构地区：[1]海南师范大学生命科学学院,海南海口571158

出　　处：《贵州农业科学》2013年第10期30-34,共5页Guizhou Agricultural Sciences

基　　金：国家自然科学基金"入侵植物银胶菊的遗传结构和生态适应性"(30960041);海南师范大学重点学科"植物生态学"(HS-1-2011-071001)

摘　　要：为了使入侵性植物银胶菊的后续试验更顺利高效,以银胶菊(Parthenium hysterophorus L.)为材料,采用L16(4)正交试验设计,对SRAP-PCR反应体系中对扩增反应影响较大的3个因素(模板DNA浓度、dNTPs浓度、引物浓度)及退火温度进行优化。结果表明:银胶菊最优SRAP-PCR反应体系25μL中含有DNA模版60ng,上下游引物各0.4μmol/L,dNTPs 0.25mmol/L,2.5μL 10×buffer(Mg2+plus 2.5μmol/L),Taq DNA聚合酶2.5U,退火温度为50℃。其中,dNTPs的用量对扩增反应影响较大,而引物用量及模板用量影响则较小。利用该体系从48对引物组合中筛选出条带丰富清晰的引物17对。In order to make the follow-on experiment successful and efficient, using P. hysterophorus as the experimental materials, the SRAP-PCR reaction system was optimized by the orthogonal experiment design Ll6 （4） with three factors, including template DNA, dNTPs concentration, primers concentration, and the annealing temperature was also optimized. The results show that the optimal SRAP-PCR reaction system is as follows： total volume 25 ttL, containing template DNA 60 ng, 0. 4 tzmol/L primers, 0.25 mmol/L dNTPs, Mg2＋ 2.5 t, mol/L, 2.5U Taq DNA polymerase,the annealing temperature is 50~C. From that, the effect of dNTPs concentration is the greatest, the primers and the template are less. 17 primer pairs were screened out from 100 primer pairs combinations based on their distinct and abundant bands by utilizing this reaction system.

关 键 词：银胶菊 SRAP-PCR 体系优化 引物筛选 

分 类 号：S452[农业科学—植物保护]