蛇皮果基因组DNA提取及其RAPD条件优化  

DNA extraction and RAPD Condition optimization for Salacca edulis Reinw

作　　者：李建光[1] 李荣生[1] 李发根[1] 尹光天[1] 杨锦昌[1] 邹文涛[1] 

机构地区：[1]中国林业科学研究院热带林业研究所,广东广州510520

出　　处：《中南林业科技大学学报》2012年第6期110-114,共5页Journal of Central South University of Forestry & Technology

基　　金：中央级公益性科研院所基本科研业务费专项项目"蛇皮果栽后遮荫和灌溉技术研究"(编号:RITFKYYW2010-02);国家林业局948引进项目"蛇皮果栽培品种及配套栽培技术引进"(编号:2007-4-11)

摘　　要：以蛇皮果顶梢嫩叶为材料,比较了CTAB法、改良CTAB法、CTAB-free法和SDS法对基因组DNA的提取效果,并对蛇皮果RAPD反应体系进行优化。结果表明,CTAB法、改良CTAB法和CTAB-free法均能提取出蛇皮果基因组DNA,其中NaCl浓度为5 m ol/L的改良CTAB法因可更有效地去除多糖及酚类物质,并能满足PCR扩增的要求而被认为是蛇皮果基因组DNA提取的最适方法;在15μL的PCR扩增体系中,Mg2+、dNTPs和Taq DNA聚合酶的最优浓度分别为1.4 mmol/L、0.12 mmol/L和0.12 U/μL;PC R反应程序的最适退火温度为39℃。By taking the tender leaves on top-shoots of Salacca edulis as tested materials, the extraction effects of CTAB, improved CTAB, CTAB-free and SDS on the genomic DNA of salak were investigated and compared. And the reaction system of salak RAPD was optimized. The results show that CTAB, improved CTAB, CTAB-free all could extracted out genomic DNA from salak, of them, the improved CTAB with NaC1 concentration of 5 mol/L could more effectively remove polysaccharides and polyphenols than the other two methods, and meet the requirement of PCR amplification, so it is the best method. The optimal concentrations of Mg2＋, dNTPs, Taq DNA polymerase and the annealing temperature are 1.4 mmol/L, 0.12 mmol/L, 0.12 U/μL and 39 ℃ respectively for the PCR progress for salak.

关 键 词：蛇皮果 DNA提取 RAPD优化 

分 类 号：S789.5[农业科学—林学]