吉林农业大学学报2011，Vol.33Issue(5)：511-517,7.DOI:22-1100/S.20110419.0835.006
2种PCR方法扩增大豆rbcS启动子5'侧翼序列比较
Comparison between Two PCR Techniques for Amplifying the 5' Flanking Sequence of the rbcS Promoter from Glycine max
摘要
Abstract
The research was conducted to compare two PCR techniques for isolating a target DNA segment flanking the known sequence. Inverse PCR(IPCR) , with two pairs of specific primers, was performed using genome DNA as the template digested with EcoRI, followed by self-ligation. TAIL - PCR, with combination of three nested specific primers and an arbitrary degenerated primer, respectively, was performed using the same genome template. 438 bp and 867 bp of specific fragments were obtained by IPCR and TAIL - PCR, Sequencing results confirmed primers and the known sequence. Comparatively, TAIL -PCR technique is very simple and reproducible and can obtain the target fragment in a shorter time. The results showed that TAIL - PCR is more efficient than IPCR in identifying the flanking sequence.关键词
反向PCR/热不对称交错PCR/大豆/rbcS启动子Key words
IPCR/ TAIL-PCR/ Glycine max/ rbcS promoter分类
生物科学引用本文复制引用
刘晓庆,丁志鑫,刘晶,崔喜艳..2种PCR方法扩增大豆rbcS启动子5'侧翼序列比较[J].吉林农业大学学报,2011,33(5):511-517,7.基金项目
国家转基因生物新品种培育重大专项(2008ZX08010-002),吉林省教育厅“十一五”科学技术研究项目[吉教科合字(2009)第61号] （2008ZX08010-002）
