Digital encoding of cellular mRNAs enabling precise and absolute gene expression measurement by single-molecule counting.

ABSTRACT: We present a new approach for the sensitive detection and accurate quantitation of messenger ribonucleic acid (mRNA) gene transcripts in single cells. First, the entire population of mRNAs is encoded with molecular barcodes during reverse transcription. After amplification of the gene targets of interest, molecular barcodes are counted by sequencing or scored on a simple hybridization detector to reveal the number of molecules in the starting sample. Since absolute quantities are measured, calibration to standards is unnecessary, and many of the relative quantitation challenges such as polymerase chain reaction (PCR) bias are avoided. We apply the method to gene expression analysis of minute sample quantities and demonstrate precise measurements with sensitivity down to sub single-cell lev

SUBMITTER: Fu GK 

PROVIDER: S-EPMC3982967 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature