RAPD的建立及优化研究

目的：建立优化的RAPD反应条件，为进一步进行遗传监控研究奠定基础。方法：按常规方法制备小鼠基因组DNA，在一定的RAPD－PCR反应条件下分别改变各组份浓度及退火温度。扩增产物在含溴化乙锭的1．5％琼脂糖凝胶中电泳，紫外透射仪上观察照相。结果：当Mg^2 浓度为1．5mmol/L,4种dNTPs各为150μmol/L,Taq酶为1．5U，引物为0．2mol/L,模板DNA为50ng,退火温度为38℃，RAPD－PCR扩增效果好。结论：在优化的条件下规范操作，RAPD的稳定性、重复性好。

Screening the optimized condition of RAPD- PCR

Objective To establish the optimized RAPD - PCR reaction conditions for further genetic monitoring study. Methods Altering the concentrations of the reaction components and the annealing temperature of RAPD - PCR, PCR products from RAPD were electrophoretically separated in agarose gels and banding profiles were visualized by ethidium bromide staining. Recults RAPD - PCR reactions were performed in duplication with SOng of genomic DNA, lSOpmol/L each dNTP, 0. 2xmol/L primer, 1. 5mmol/L magnesium chloride, I.. 5 units of Taq polymerase, annealing at 38t . Under these conditions RAPD method is stable and reproducible. Conclusion The optimized RAPD - PCR reaction conditions were established in this study.