大规模平行测序技术(MPSS)研究进展

A Novel Gene Identification Approach: Massively Parallel Signature Sequencing

大规模平行测序技术 (massivelyparallelsignaturesequencing ,MPSS)是以DNA测序为基础的大规模高通量基因分析新技术 ,通过标签库的建立、微珠与标签的连接、酶切连接反应和生物信息分析等步骤 ,获得基因表达序列 .MPSS具有能测定表达水平较低、差异较小的基因 ,不必预先知道基因的序列 ,自动化和高通量等特点 。

Massively parallel signature sequencing, MPSS, is an open platform that reveals the expression level of virtually all genes expressed in a sample by counting the number of individual mRNA molecules produced from each gene. The MPSS process involves cloning each mRNA molecule onto the surface of a 5 mum bead. The DNA combitag sequence is attached to a fragment of cDNA. The cDNA library is hybridized to beads. After hybridization, each of the beads displays amplified copies of one and only one starting mRNA molecule. MPSS has a routine sensitivity of a few molecules of mRNA per cell and the results are in a digital format that simplifies data management and analysis. MPSS results will be particularly. useful for generating the type of complete data sets that will help to identify the functionally important genes in the sample of interest.