秦巴山区野生山丹百合DNA提取与RAPD反应体系建立  

A Study on DNA Extraction and Establishment of RAPD Reaction System in Lilium pumilum DC. Fisch from Qinba Mountainous Area

作　　者：钟海丰[1] 张延龙[1] 牛立新[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农业学报》2008年第3期285-289,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(30571523)

摘　　要：以秦巴山区野生山丹百合幼嫩叶片为材料,采用常规CTAB法提取百合基因组DNA,得到的DNA基本能满足RAPD-PCR分析。通过单因素逐一递进优化,得到在20μL的PCR反应体系中,野生山丹百合RAPD分析的最佳反应体系为:40 ng的模板DNA,0.4μmol/L随机引物,2.25 mmol/L Mg2+,0.2 mmol/LdNTPs,Taq DNA聚合酶1.5U,1×buffer缓冲液,ddH2O补足20μL。RAPD扩增反应程序为:94℃预变性4 min,94℃30 s,37℃45 s,72℃90 s,35个循环,最后72℃10 min,4℃保存。该反应体系具有较好的稳定性及可重复性。The materials for the DNA extraction which used the method of CTAB were some samples of leaves which taken from Lilium pumilum DC.Fisch that came from the Qinba mountainous area.And the optimized reaction system and program of RAPD for were as follows.The reactions were performed in a volume of 20 μL,which containing 40ng DNA template,0.4 μmoL random primer,2.25 mmol/L Mg^2＋,0.2 mmol/L dNTPs,1.5U Taq DNA polymerase,1×buffer solution.The RAPD reaction was programmed by 1 cycle for 4min at 94℃,followed by 35 cycles for 30 s at 94℃,45 s at 37℃,and 90 s at 72℃ and finally,by 1 cycle for 10 min at 72℃,storing at 4℃.The reaction system was well reproducible and highly reliable,and it could be effectively for RAPD analysis in Lilium pumilum DC.Fisch.

关 键 词：秦巴山区 山丹百合 DNA提取 RAPD反应体系建立 

分 类 号：S682.29[农业科学—观赏园艺]