A NOVEL METHOD FOR GENE EXPRESSION PROFILING: BIOLUMINOMETRIC ASSAY COUPLED WITH A MODIFIED REVERSE-TRANSCRIPTION PCR (BA mRT-PCR)
With the complete genome sequences of many organisms being reported, methods to carry out systematic collection of gene expression data will play an important role in the future of basic research. Here we describe a novel method for gene expression profiling based on pyrosequencing. This method includes four steps: (1) reverse-transcription PCR (RT-PCR) by using an anchor primer consisting of a poly (T) sequence in 3 -end. a universal sequence in 5 -end, and a source-specific sequence in the middle; (2) pooling cDNAs from different sources; (3) amplification with a universal primer and a gene specific primer; (4) pyrosequencing to detect the relative peak intensity between each of source-specific bases. The signal ratio reflects the proportion of the amount of mRNAs between different sources. For the comparison, real-time PCR (RT-PCR) was also carried out. The results from two different methods were almost the same, indicating that our new method was promising in quantitatively comparing gene expression level between different sources.
Gene expression modified reverse-transcription pyrosequencing
Xiaodan Zhang Haiping Wu Guohua Zhou
Huadong Research Institute for Medicine and Biotechnics, Nanjing 210096, China Huadong Research Institute for Medicine and Biotechnics, Nanjing 210096, China;Medical School, Nanji
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
62-66
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）