摘要： 为快速建立优化的芍药SRAP扩增反应体系,应用L25(56)正交表,研究了Taq、Mg2+、随机引物、dNTPs和DNA模板5种反应组分的浓度变化对SRAP扩增结果的影响,直观分析和稳定性检测结果表明:正交设计可高效建立优化稳定的芍药SRAP反应体系;用该法建立的芍药SRAP-PCR优化反应体系为:25μL反应体系中含10×PCR Buffer(100 mmol/L Tris-HCl pH 8.3,500 mmol/L KCl)2.5μL,MgCl22.5 mmol/L,dNTP各0.25 mmol/L,引物各0.3μmol/L,TaqDNA聚合酶1 U,DNA模板120 ng。
关键词:
芍药,
SRAP-PCR,
正交设计
Abstract: To construct the optimum SRAP reaction system of Paeonia lactif lora Pall., orthogonal experimental design L25(56) was used.And the comprehensive effect of five levels of five factors ( Taq polymerase,Mg2+, dNTP, primer and DNA template) was assessed.According to the amplificat ion results, treatment 18 was used for the subsequent detection for stability with other 5 cultivars. The final results showed that by orthogonal design, an optimal and stable SRAPPCR system for herbaceous peonies was quickly obtained: 2.5mmol/ L Mg2+, 0.25 mmol/L dNTP, 0.3 mol/L primer,1 U Taq polymerase, 120 ng DNA template within total 25 ??L reaction solution.
Key words:
Paeonia lactif lora Pall. ,
SRAP??PCR,
Orthogonal design
中图分类号:
孙宪芝, 王晓菡, 马燕, 郭先锋, 于晓艳. 应用正交设计建立芍药的SRAP反应体系[J]. 华北农学报, 2008, 23(S2): 198-200. doi: 10.7668/hbnxb.2008.S2.045.
SUN Xian-zhi, WANG Xiao-han, MA Yan, GUO Xian-feng, YU Xiao-yan. Establishment of the SRAP Reaction System of Paeonia lactiflora Pall.by Orthogonal Design[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(S2): 198-200. doi: 10.7668/hbnxb.2008.S2.045.