江汉大学学报：自然科学版2012，Vol.40Issue(5)：71-75,5.
串联表达载体中短重复序列（BmKb1）n模板的PCR效率及其优化条件研究
PCR Efficiency of Short Repetitive Sequences (BmKb1)n in Tandem Expression Vector and Optimization of Its PCR Conditions
摘要
Abstract
After acquiring BmKbl gene by Overlap PCR, pGEX-6p-1/BmKbl vector was constructed. The series recombinant pGEX-6p-1/(BmKbl)n (n = 2, 4, 6, 8, 10) plasmids were build with isocaudamer technology. With the template of pGEX-6p-1/(BmKbl)n standard PCR procedure was used to study PCR efficiency of the short tandem repeats; The BmKbl octoploid template was used to study the effects of the number of PCR cycles, the final concentration of primers and the annealing temperature on PCR efficiency and specificity of the short sequence re- peat, respectively. The results showed that: with the number of tandem repeat sequences increas- ing, the number of non-specific bands will increase, and the content of specific bands will de- crease; Amplifying BmKbl gene tandem by PCR in the 18-20 cycle numbers, with the primer final concentration of 0.05-0.1μM, as well as at higher annealing temperature of 60-62℃, the relatively high yield and high the specificity PCR fragments of BmKbl gene concatemer will be available. This study will provide a technology basis for the cloning, PCR screening, sequencing of tandem expression vector and genome STRs.关键词
重复序列/PCR/BmKb1/循环数Key words
tandem repeats/PCR/BmKbl/cycle number分类
生物科学引用本文复制引用
范丽梅,李正,余腾,刘燕群,罗锋..串联表达载体中短重复序列（BmKb1）n模板的PCR效率及其优化条件研究[J].江汉大学学报：自然科学版,2012,40(5):71-75,5.基金项目
武汉市科技局青年科技晨光计划项目 （）
武汉市市属高等学校科学研究重点项目 （）
湖北省高等学校优秀中青年科技创新团队项目 （）
