[Preparation of prokaryotic cDNA for high-throughput transcriptome analysis]

Bioorg Khim. Nov-Dec 2011;37(6):854-7. doi: 10.1134/s1068162011060045.
[Article in Russian]

Abstract

High contents of non-coding RNA in total bacteria RNA complicates considerably transcriptome analysis using standard approaches like high-throughput sequencing, gene expression profiles, subtractive hybridization. We suggest a procedure of preparation of bacterial cDNA for transcriptomics that includes rRNA and tRNA depletion with preservation of relative abundance of coding sequences. The method is based on the second order hybridization kinetics and unique properties of Kanchatka crab duplex-specific nuclease. The method efficacy was demonstrated on a model experiments.

Publication types

  • English Abstract

MeSH terms

  • DNA, Complementary / isolation & purification*
  • Gene Expression Profiling*
  • Genome, Bacterial
  • High-Throughput Screening Assays*
  • Prokaryotic Cells / chemistry
  • RNA, Untranslated / chemistry

Substances

  • DNA, Complementary
  • RNA, Untranslated