Molecular indexing enables quantitative targeted RNA sequencing and reveals poor efficiencies in standard library preparations.

ABSTRACT: We present a simple molecular indexing method for quantitative targeted RNA sequencing, in which mRNAs of interest are selectively captured from complex cDNA libraries and sequenced to determine their absolute concentrations. cDNA fragments are individually labeled so that each molecule can be tracked from the original sample through the library preparation and sequencing process. Multiple copies of cDNA fragments of identical sequence become distinct through labeling, and replicate clones created during PCR amplification steps can be identified and assigned to their distinct parent molecules. Selective capture enables efficient use of sequencing for deep sampling and for the absolute quantitation of rare or transient transcripts that would otherwise escape detection by standard sequencing

SUBMITTER: Fu GK 

PROVIDER: S-EPMC3918775 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature