正交试验与单因子双重试验法优化玉米PCR反应体系  

Optimization of PCR Reaction System of Corn by Orthogonal Test and Single Factor Double Test

作　　者：江常胜 朱洪海 钟永丽 刘明晓 东方 JIANG Changsheng;ZHU Honghai;ZHONG Yongli

机构地区：[1]中禾生物种业集团有限公司,广东深圳518000

出　　处：《智慧农业导刊》2022年第21期14-17,共4页JOURNAL OF SMART AGRICULTURE

基　　金：深圳市科技计划项目(JSGG20180504165849329)。

摘　　要：为探索出玉米PCR反应的最佳体系,解决玉米分子育种工作中基因序列扩增问题,给育种工作提供便利,该文试验以玉米幼嫩叶片为材料,采用正交试验设计法对影响玉米PCR反应的引物用量、模板和模板质量浓度等因素进行3因素4水平优化;对退火温度进行单因子8水平试验优化,通过试验探索出玉米10μLPCR的最佳反应体系为5μL2×EasyTaq~?PCR SuperMix(含EasyTaq~?DNA Polymerase、dNTPs)、引物0.25μL和1μL40 ng/μL模板DNA,退火温度56.6℃。In order to explore the best system of corn PCR reaction,solve the problem of gene sequence amplification in corn molecular breeding,and provide convenience for corn breeding,with young corn leaves as materials,the amount of primers,template and template mass concentration affecting corn PCR reaction were optimized by orthogonal design.The an-nealing temperature was optimized by single factor 8-level experiment.The optimum reaction system of corn 10μLPCR was found as follows 5μL 2×EasyTaqR PCR SuperMix(including EasyTaqR DNA Polymerase and dNTPs),primers 0.25μL,1μL40 ng/μL template DNA,and the annealing temperature is 56.6℃.

关 键 词：分子育种 玉米 PCR 正交试验设计 单因子试验 体系优化 

分 类 号：S-3[农业科学]