改良一步法分离变形链球菌RNA  

Modified one-step extraction of RNA from Streptococcus mutans

作　　者：郭青玉[1] 吴军正[1] 肖刚[2] 李睿[2] 

机构地区：[1]第四军医大学秦都口腔医院口腔生物教研室,西安710032 [2]西安交通大学口腔医院口腔内科,西安710004

出　　处：《口腔医学》2007年第1期32-34,共3页Stomatology

摘　　要：目的快速、高效分离高质量变形链球菌RNA。方法溶菌酶处理变形链球菌后加入10%SDS,再经沸水浴1-2min可快速裂解细菌;裂解产物以一步法分离RNA。琼脂糖凝胶电泳及紫外分光光度计观察和测量RNA完整性及浓度。结果与其他方法比较,该法所获RNA产量高,几乎达30μgRNA/109细胞且无DNA污染。反转录（RT）-PCR证实mRNA的完整性好。结论改良一步法简便、经济、稳定,所得RNA可用于后续研究。Objective To isolate high quality RNA from S. mutans efficiently. Method We used a modified one-step method to extract RNA. Rapid lysis of S. mutans was obtained with lysozyme, followed by 10%SDS treatment and boiled water bath for 1-2 mins. Lysates were successfully used for one extraction of RNA. RNA integrity and yield were assessed by agarous gel eleetrophoresis and the absorbanee at 260 nm.Results Compared with other procedures, this method yielded a higher level of RNA, almost 30 μg/10^9 cells. DNA was not co-recovered. The presence of intact mRNA within the total RNA fraction was demonstrated by RT-PCR. Conclusions It is concluded that the improved one-step method is a reproducible, economic and simple way to permit rapid isolation with high yield of intact RNA for subsequent analysis.

关 键 词：RNA 变形链球菌 细菌裂解 

分 类 号：R349[医药卫生—基础医学]