Construction of sized eukaryotic cDNA libraries using low input of total environmental metatranscriptomic RNA.

ABSTRACT:

Background

Construction of high quality cDNA libraries from the usually low amounts of eukaryotic mRNA extracted from environmental samples is essential in functional metatranscriptomics for the selection of functional, full-length genes encoding proteins of interest. Many of the inserts in libraries constructed by standard methods are represented by truncated cDNAs due to premature stoppage of reverse transcriptase activity and preferential cloning of short cDNAs.

Results

We report here a simple and cost effective technique for preparation of sized eukaryotic cDNA libraries from as low as three microgram of total soil RNA dominated by ribosomal and bacterial RNA. cDNAs synthesized by a template switching approach were size-fractionated by two dimensional agarose gel electro

SUBMITTER: Yadav RK 

PROVIDER: S-EPMC4170940 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature