黄瓜AFLP反应体系的优化  

Optimization of AFLP Reaction System in Cucumber

作　　者：张素勤[1] 顾兴芳[1] 张圣平[1] 王晓武[1] 邹志荣[2] 

机构地区：[1]中国农业科学院蔬菜花卉研究所 [2]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271-273,共3页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家863资助项目(2001AA2411212002AA244012003AA207120);农业部重点实验室项目

摘　　要：对影响AFLP(Amplified fragment length polymorphism)扩增的关键因素进行了摸索,以期获得清晰稳定的黄瓜AFLP反应体系。结果发现,最佳的预扩增体系为:20反应体系中包含0.5 UTaqDNA poly-merase、2.0μL dNTP(2 mmol/L)、1.2μL 25 mmol/L Mg2+、0.6μL 50 ng/μL每种引物、1×PCR Buffer和4μL模板DNA;最优的选择性扩增体系为20μL反应体系中含有1.0 UTaqDNA polymerase、2.0μL dNTP(2 mmol/L)、1.0μL 25 mmol/L Mg2+、1.0μL 50 ng/μL每种引物、3.0μL稀释40倍的预扩增产物和1×PCR Buffer。体系优化后,扩增产物稳定,条带十分清晰,无背景干扰。The key factors influencing AFLP（amplified fragment length polymorphism） amplification were researched in order to establish a highly stable and optimized AFLP reaction system in cucumber in the present study.The results showed that the optical pre-amplification system was 20 μL reaction mix containing 0.5 unit of Taq DNA polymerase,2.0 μL of dNTP（2 mmol/L）,1.2 μL of 25 mmol/L Mg^2＋,0.6 μL of each primer（50 ng/μL）,4 μL of template DNA and 1× buffer,the optimized selective amplification protocol was 20 μL reaction mix containing 1.0 unit of Taq DNA polymerase,2.0 μL of dNTP（2 mmol/L）,1.0 μL of 25 mmol/L Mg^2＋,1.0 μL of each primer（50 ng/μL）,3.0 μL of 40× diluted product of pre-amplification and 1× buffer.Amplified bands were clear and stable without disturbed background after the optimization of AFLP reaction system.

关 键 词：黄瓜 AFLP 反应体系 优化 

分 类 号：S642.2[农业科学—蔬菜学]