贵州医科大学学报

2008, No.135(06) 635-639

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家蝇相关序列扩增多态性扩增体系的建立与优化
Establishment and Optimization of SRAP-PCR Amplification System for Housefly

刘红美,方小波,郑勤妮,胡仕明,吴建伟
LIU Hongmei1,FANG Xiaobo1,ZHENG Qinni1,HU Shiming1,WU Jianwei2 (1. Department of Medical Biotechnology

摘要(Abstract):

目的:建立和优化家蝇的相关序列扩增多态性(SRAP-PCR)扩增体系。方法:以驯养家蝇为材料,通过单因子实验分别研究了模板DNA、Mg2+、dNTPs、引物浓度及Taq DNA聚合酶用量对家蝇SRAP分子标记扩增体系的影响,并对扩增体系进行了优化。结果:优化后的反应体系总体积为30μl,含50ng模板DNA、1.5mmol/LMg2+、0.20mmol/L dNTPs、0.15μmol/L引物和0.50U Taq DNA聚合酶。结论:家蝇SRAP-PCR扩增优化系统的建立,为今后利用SRAP技术进行家蝇遗传多样性分析、基因定位奠定了基础。
Objective:To establish and optimize SRAP-PCR reaction system of housefly. Methods:Effects of concentrations of template DNA,Mg2+,dNTPs,primers and amount of Taq DNA polymerase on SRAP molecular marker amplification system of Musca domestica L were studied in monofactorial experiment,and the amplification system was optimized. Results:Total volume of the optimal system was 30μl containing 50 ng of template DNA,1.5mmol/ L of Mg2+,0. 20 mmol/ L of dNTPs,0. 15μmol/ L of primers and 0. 50 U of Taq DNA polymerase. Conclusion:This system provides a useful tool for analysis of genetic diversity and gene location of Musca domestica L.

关键词(KeyWords): 蝇科;随机扩增多态DNA技术;遗传标记
Muscidae; random amplified polymorphic DNA technique; genetic markers

Abstract:

Keywords:

基金项目(Foundation): 贵州省优秀科技教育人才省长基金(S2004-17);; 贵州省高层次人才科研条件特助经费(Q2005-4);; 贵阳医学院博士启动基金(C2005-6)

作者(Author): 刘红美,方小波,郑勤妮,胡仕明,吴建伟
LIU Hongmei1,FANG Xiaobo1,ZHENG Qinni1,HU Shiming1,WU Jianwei2 (1. Department of Medical Biotechnology

DOI: 10.19367/j.cnki.1000-2707.2008.06.020

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