Removing PCR for the elimination of undesired DNA fragments cycle by cycle

文献类型： 外文期刊

第一作者： Huan, Jiaojiao

作者： Huan, Jiaojiao;Wan, Kangkang;Dong, Wubei;Huan, Jiaojiao;Wan, Kangkang;Dong, Wubei;Liu, Yunjun;Wang, Guoying;Liu, Yunjun;Wang, Guoying

作者机构：

期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）

ISSN： 2045-2322

年卷期： 2013 年 3 卷

页码：

收录情况： SCI

摘要： A novel removing polymerase chain reaction (R-PCR) technique was developed, which can eliminate undesired genes, cycle by cycle, with efficiencies of 60.9% (cDNAs), 73.6% (genomic DNAs), and, similar to 100% (four DNA fragments were tested). Major components of the R-PCR include drivers, a thermostable restriction enzyme - ApeKI, and a poly(dA) adapter with mismatched restriction enzyme recognition sites. Drivers were generated from the undesired genes. In each cycle of R-PCR, drivers anneal to complementary sequences and allow extension by Taq DNA polymerase. Thus, ApeKI restriction sites in the undesired genes are recovered, and adapters of these undesired DNA fragments are removed. Using R-PCR, we isolated maize upregulated defense-responsive genes and Blumeria graminis specialized genes, including key pathogenesis-related effectors. Our results show that after the R-PCR reaction, most undesired genes, including very abundant genes, became undetectable. The R-PCR is an easy and cost-efficient method to eliminate undesired genes and clone desired genes.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序