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| 线虫PCR:直接针对秀丽线虫进行PCR扩增基因组DNA的新方法 | |
| 引用本文： | 李媛,任长虹,石锦平,李伟光,刘虎岐,张成岗. 线虫PCR:直接针对秀丽线虫进行PCR扩增基因组DNA的新方法[J]. 生物技术通讯, 2011, 22(1): 66-70. DOI: 10.3969/j.issn.1009-0002.2011.01.016 |
| 作者姓名： | 李媛 任长虹 石锦平 李伟光 刘虎岐 张成岗 |
| 作者单位： | 1. 西北农林科技大学,生命科学学院,陕西,杨凌,712100;军事医学科学院,放射与辐射医学研究所,蛋白质组学国家重点实验室,北京,100850 2. 军事医学科学院,放射与辐射医学研究所,蛋白质组学国家重点实验室,北京,100850 3. 西北农林科技大学,生命科学学院,陕西,杨凌,712100 |
| 基金项目： | 国家重点基础研究发展计划(2006CB504100); 国家自然科学基金(30900862,30800196,30771230); 国家科技重大专项(2009ZX09503-002,2009ZX09301-002,2009ZX09103-616) |
| 摘 要： | 目的:直接针对秀丽线虫进行PCR反应,以便快速扩增基因组DNA,从而提高钓取目的基因和鉴定基因组是否发生突变的效率.方法:根据生物信息学分析,针对不同基因设计单重或多重PCR引物;在不含砌DNA聚合酶的PCR反应体系中加入蛋白酶K消化秀丽线虫染色体中的组蛋白,然后加入Taq酶,直接针对野生型或突变型秀丽线虫个体进行PC... |
| 关 键 词： | 秀丽线虫 线虫PCR 多重PCR 基因扩增 突变株系鉴定 |
A New Method to Amplify the Genomic DNA by Using PCR on the Caenorhabditis elegans Directly |
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| LI Yuan,REN Chang-Hong,SHI Jin-Ping,LI Wei-Guang,LIU Hu-Qi,ZHANG Cheng-Gang. A New Method to Amplify the Genomic DNA by Using PCR on the Caenorhabditis elegans Directly[J]. Letters in Biotechnology, 2011, 22(1): 66-70. DOI: 10.3969/j.issn.1009-0002.2011.01.016 | |
| Authors: | LI Yuan REN Chang-Hong SHI Jin-Ping LI Wei-Guang LIU Hu-Qi ZHANG Cheng-Gang |
| Affiliation: | LI Yuan1,2,REN Chang-Hong2,SHI Jin-Ping2,LI Wei-Guang2,LIU Hu-Qi1,ZHANG Cheng-Gang1,2 1.College of Life Science,Northwest Agriculture and Forest University,Yangling 712100,2.Beijing Institute of Radiation Medicine,State Key Laboratory of Proteomics,Beijing 100850,China |
| Abstract: | Objective:In order to improve the efficiency of target gene amplification and mutant identification,the Caenorhabditis elegans was used directly in the PCR.Methods:According to bioinformatics analysis,primers for single or multiplex PCR were designed.Firstly,the proteinase K and the C.elegans(either wild type or mutants) were added into the PCR system without Taq DNA polymerase.Secondly,proteinase K was inactivated by heating.Finally,the Taq polymerase was added and the PCR procedure started to amplify the ... |
| Keywords: | Caenorhabditis elegans C.elegans PCR multiplex PCR gene amplification mutant identification |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |