花生SRAP-PCR反应体系的正交设计优化  

Optimization for SRAP-PCR System of Peanut Based on Orthogonal Design

作　　者：邹小云[1] 邹晓芬[1] 陈伦林[1] 李书宇[1] 张建模[1] 陈志才[1] 宋来强[1] 

机构地区：[1]江西省农业科学院作物研究所,江西省农业科学院油料作物重点实验室,南昌330200

出　　处：《分子植物育种》2010年第4期822-826,共5页Molecular Plant Breeding

基　　金：江西省科技支撑计划项目;江西省农业科学院产业技术支撑体系建设项目资助

摘　　要：本研究利用正交设计L16(45)对花生SRAP-PCR反应体系的5因素(引物,Taq酶,Mg2+,模板DNA和dNTP)在4水平上进行优化试验,结果表明,各因素的不同水平对PCR反应结果都有显著的影响,其中引物浓度的影响最大;最佳反应体系20μL包含:引物0.4μmol/L,Taq酶1U,Mg2+2.5mmol/L,模板DNA30ng,dNTP0.2mmol/L,不足部分以ddH2O补足。这一体系的建立为今后利用SRAP标记技术对花生进行分子遗传学基础研究提供了有力的支持。We employed the orthogonal design to optimize SRAP-PCR system on peanut in four levels of five factors,primer,Taq DNA polymerase,Mg^2＋,DNA template,and dNTP,respectively.In this research the results showed that there were significant effects under the different combination of four levels and five factor,and than the primer concentration was the most dominant factor affected on the result of PCR.The orthogonal experiment design was a practicable method and in a total volume of 20 μL SRAP-PCR system,it contains 0.4 μmol/L primer,1 U Taq DNA polymerase,2.5 mmol/L Mg^2＋,30 ng DNA template,0.2 mmol/L dNTP and the rest of them was ddH2O.This system would provide a powerfull support for molecular genetics research by using SRAP technique of peanut in the future.

关 键 词：花生 SRAP 正交设计 体系优化 

分 类 号：S565.2[农业科学—作物学]