不同方法鉴定差异显示技术RAP-PCR得到差异基因的比较

Comparison of Three Methods for Confirmation of Differential Fragments from RAP-PCR

目的:评价用RAP-PCR法从测序胶上得到深海细菌(Shewanella piezotolerance WP3)差异片段验证的最佳方法。方法:用RT-PCR,RNA点杂交,荧光实时定量PCR对不同盐浓度的差异片段进行对比实验。结果:3种方法均能检测到片段的差异表达:RT-PCR灵敏性最强,RNA点杂交需要的RNA量最大,荧光实时定量PCR精确性最高。结论:在实验条件允许下,用荧光实时定量PCR对较少片段的验证较好。

Objective： To test the best method for confirming differential fragments from the sequencing gels. Methods： RNA samples of the cultures from the different salt concentrations were taken as a model. RT-PCR, RNA dot hybridization and real-time PCR were used for confirmation of differential fragments. Results： Three methods could produce good results. RT-PCR method had high sensibility, RNA dot hybridization needed huge quantitative RNA sample and the realtime PCR had high accuracy. Conclusion： Real-time PCR which can quantify the change folds can be a better choice.