3种mRNA检测方法比较

Comparing three methods for detection of mRNA expressions

目的 :比较 3种检测mRNA表达的方法。方法 :采用Nothernblot,RT PCR ,原位杂交法检测基因mRNA的表达。结果及结论 :在使用Nothernblot杂交进行定量分析时 ,关键因素有 :①RNA的质量 ;②探针标记效率 ;③洗膜的控制。RT PCR检测很难用于定量分析。原位杂交法可进行细胞内mRNA的定位分析但不能确定mRNA的绝对含量。将这些方法结合使用 ,可全面了解基因mRNA的表达。

Aim:To compare three methods for detecting expressions of a gene mRNA Methods:Northern blot, RT PCR, in situ hybridization were used for examining expressions of a gene mRNA Results and Conclusion:The key issues in using Nothern blot for quantitative analysis are high quality RNA, efficiency of marked probe and control of washing membrane RT PCR is no suitable for quantitative analysis In situ hybridization may be used for the located analysis of mRNA in a cell, but no confirmation for absolute content of mRNA in a cell Overall information of mRNA expressions may be obtained by combination of these methods