甘蓝SRAP-PCR反应体系的建立  

Establishment and Optimization of an SRAP-PCR Detection System of Cabbage

作　　者：杨仕春[1] 李成琼[1] 司军[1] 任雪松[1] 宋洪元[1] 

机构地区：[1]西南大学园艺园林学院,重庆市蔬菜学重点实验室,南方山地园艺学教育部重点实验室,重庆400716

出　　处：《西南大学学报（自然科学版）》2013年第2期11-14,共4页Journal of Southwest University(Natural Science Edition)

基　　金：国家科学支撑计划项目(2008BADB1B02);重庆市农作物良种创新工程资助项目(CSTC,2010AA1023);中央高校基本科研业务费资金资助项目(XDJK2010C068)基金资助

摘　　要：以甘蓝自交系南宝为试验材料,采用L16(45)正交设计,对影响甘蓝SRAP的5个因素(模板DNA浓度、Mg2+浓度、dNTPs浓度、引物浓度、Taq酶)在4个水平上进行优化试验,建立了可扩增多态性高、重复性好、稳定性强、带型清晰的最优反应体系(10μL):模板DNA 40ng、MgCl22.50mmol/L、dNTPs 0.20mmol/L、Primer0.30μmol/L、Taq酶0.40U,该体系能满足甘蓝基因组SRAP扩增的要求.In this study, the cabbage inbred line NANBAO was adopted as the experiment material and the orthogonal design of five factors （DNA template concentration, Mg2＋ concentration, dNTPs concentra- tion, primer concentration and Taq DNA polymerase） at four levels was used to establish an optimized SRAP-PCR system of cabbage. The optimal concentrations of the five factors in the 10 μL SRAP system were DNA template 40 ng, MgC12 2.50 mmol/L, dNTPs 0.20 mmol/L, primer 0.30 μmol/L and Taq DNA polymerase 0.40 U. The system thus established was reliable, stable, reproductive and legible and could meet the demands for genome SRAP amplification in cabbage.

关 键 词：甘蓝 SRAP-PCR 体系 建立 

分 类 号：S635.1[农业科学—蔬菜学] Q784[农业科学—园艺学]