【摘要】 以芸芥9370为材料,在SDS法的基础上,通过改变离心时间,操作步骤,建立了一种较快速提取芸芥基因组DNA的方法。使用美国伯乐公司生产的Mycycler TM thermal cycler扩增仪,对影响RAPD扩增的因素进行了比较研究,确定了模板、10×buffer(主要含Mg2+)、、dNTPs、引物和Taq DNA聚合酶的适宜量和最佳的循环次数。试验结果表明在25μL体系中使用18.5μL ddH2O,2μL 10×buffer(主要含Mg2+),2μLdNTPs,1μL引物,0.5μL TaqDNA聚合酶,1μL模板;反应条件为94℃预变性4 min,然后进行94℃50 s,38℃退火1 min 20 s,72℃延伸2 min,40个循环后,再在72℃延伸10 min,芸芥RAPD扩增效果较好。更多还原

【Abstract】 A simple and handy method to extract genomic DNA from the leaves of E.sativa Mill was established by making use of SDS method and changing centrifugal time,process.The factors affected the RAPD pattern were studied.A stable RAPD reaction system for E.sativa Mill was set up.This system was 25 μL total volume contained 18.5 μL ddH2O,2 μL 10×buffer(Mg2+),1 μL Primer,0.5 μL TaqDNA polymerase,1 μL Template DNA,2 μL dNTPs.After pre-denaturing at 94℃ for 5 min,under the condition of denaturing at 94℃ for 50 seconds,annealing at 38℃ for 1 min for 20 seconds,extension at 72℃ for 90 seconds,amplifying for 40 cycles,and extension at 72℃ for 10 min,the results of amplification was good for RAPD research in E.sativa Mill.更多还原