A method for the construction of equalized directional cDNA libraries from hydrolyzed total RNA.

ABSTRACT:

Background

The transcribed sequences of a cell, the transcriptome, represent the trans-acting fraction of the genetic information, yet eukaryotic cDNA libraries are typically made from only the poly-adenylated fraction. The non-coding or translated but non-polyadenylated RNAs are therefore not represented. The goal of this study was to develop a method that would more completely represent the transcriptome in a useful format, avoiding over-representation of some of the abundant, but low-complexity non-translated transcripts.

Results

We developed a combination of self-subtraction and directional cloning procedures for this purpose. Libraries were prepared from partially degraded (hydrolyzed) total RNA from three different species. A restriction endonuclease site was added to

SUBMITTER: Davis C 

PROVIDER: S-EPMC2134933 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature