白僵菌RAPD-PCR反应体系优化  

RAPD-PCR Reaction System for Beauveria bassiana

作　　者：魏晓鹏[1] 李会平[1] 黄大庄[1] 唐秀光[1] 

机构地区：[1]河北农业大学,保定071000

出　　处：《东北林业大学学报》2012年第1期73-75,共3页Journal of Northeast Forestry University

基　　金：河北省自然科学基金项目(C2011204041)

摘　　要：为了建立一套适宜于白僵菌(Beauveria bassiana)退化研究的RAPD-PCR反应体系及反应程序,通过采用L16(45)正交试验及退火温度和循环次数的单因素优化对反应体系中的各因素进行优化组合。结果表明:20μL PCR反应体系及反应程序中各因素优化组合为,10×Buffer 2μL,MgCl2(25 mmol/L)2.4μL,4种dNTP(各2.5mmol/L)0.8μL,随机引物(10μmol/L)1.4μL,TaqDNA聚合酶(5 U/μL)0.4μL,模板DNA(10 mg/L)1μL。反应条件为,94℃预变性2 min,94℃变性30 s,38℃退火40 s,72℃延伸1 min,循环次数40次,72℃延伸5 min。An experiment was conducted to establish an optimal PCR （polymerase chain reaction） reaction system and procedure for the degradation of Beauveria bassiana. Single factor test and L16（45 ） orthogonal experiment were used to optimize the combination of factors for the reaction system. The optimum factor combination was obtained with 20 μL reaction volume containing 2μL 10xBuffer, 2.4μL M b, C12 （25 retool/L）, 0.8 μL four types of dNTPs （each 2.5 mmol/L）, 1.4 μL random primer （ 10 μmol/L）, 0.4μL Taq polymerase （5 U/μL）, and 1μL template DNA （ 10 mg/L）. Reaction conditions were as follows ： predenaturing at 94 degrees C for 2 min, followed by 40 cycles of denaturing at 94 degrees C for 30 s, annealing at 38 degrees C for40 s, extension at 72 degrees C for 1 min, and final extension at 72 degrees C for5 min.

关 键 词：白僵菌 RAPD—PCR 正交组合 

分 类 号：Q939.53[生物学—微生物学]