摘要
以意大利蜜蜂为材料,研究了蜜蜂DNA的提取以及对RAPD分析的影响因素,包括模板浓度、Mg2+、dNTP和引物,建立了适于蜜蜂RAPD分析的PCR反应体系,即20μL反应体系中,包括10mmol/LTris-HCl(pH8.3)、50mmol/LKCl、20～60ngDNA、3.0mol/LMgCl2、0.2mmol/LdNTP、0.5μmol/L随机引物和1.5unitTaq聚合酶。扩增程序为:94℃预变性5min,94℃变性1min,36℃退火1min,72℃延伸2min,40个循环;最后在72℃延伸10min。
DNA extraction and factors influencing RAPD analysis,including the concentration of DNA template,Mg2+,dNTP and primers,in bees were studied. An optimal condition of amplification for RAPD analysis in bees was established: in 20 μL reaction solution,containing 10 mmol/L Tris-HCl(pH8.3),50 mmol/L KCl,20～60 ng DNA template,3.0 mol/L MgCl2,0.2 mmol/L dNTP,0.5 μmol/L random primers and 1.5 U polymerase. The amplification program was devised: 94℃ for 5 min, denaturing at 94℃ for 1 min, primers annealing at 36℃ for 1 min, extension at 72℃ for 2 min ,40 cycles; at last extension at 72℃ for 10 min.
出处
《江西农业大学学报》
CAS
CSCD
2003年第5期783-786,共4页
Acta Agriculturae Universitatis Jiangxiensis
基金
国家自然科学基金资助项目(30160068)
