摘要
目的为了建立RAPD-PCR鉴定分枝杆菌的方法,同时了解其影响因素。方法采用不同引物及其浓度、不同DNA提取方法及其他反应条件进行随机引物PCR。结果分枝杆菌DNA提取后,-20℃30天内结果无影响。扩增的退火温度较低,对反应条件高度依赖。在研究中发现,实验条件的变化,可产生缺带现象,此时需要重复2次,如果3次扩增的指纹图谱相同,则可以确定。结论RAPD-PCR扩增条件的优化组合是获得稳定、可靠的分枝杆菌DNA指纹的关键。
Objective To establish a method to identy mycobacteria using randomly amplified polymorphic DNA (RAPD 鈥?PCR). Methods Template DNA was treated and the concentration of primer and anneal temperature were adjusted and established. Results Purified DNA stored at 鈥?20oC was stable. The banding patterns obtained by RAPD鈥擯CR were not reproducible. Conclusion Optimization of the reaction system of RAPD 鈥擯CR is the key to stablize DNA findgerprint patterns of mycobacteria.
出处
《临床肺科杂志》
2005年第2期154-157,共4页
Journal of Clinical Pulmonary Medicine
基金
广东省医学科研基金资助项目(20004042)
