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Russian Journal of Bioorganic Chemistry

, Volume 37, Issue 6, pp 775–778 | Cite as

Preparation of prokaryotic cDNA for full-scale transcriptome analysis

  • E. A. Bogdanova
  • I. A. Shagina
  • Yu. G. Yanushevich
  • L. L. Vagner
  • S. A. Lukyanov
  • D. A. Shagin
Letter To The Editor
  • 109 Downloads

Abstract

A high content of noncoding RNA in the total bacterial RNA significantly impedes analysis of their transcriptome using standard approaches, i.e., full-scale sequencing, analysis of gene expression profiles, and subtractive hybridization. A method of preparation of bacterial cDNA for transcriptome analysis is proposed that includes a depletion of the excess of rRNA and tRNA copies with the preservation of the relative abundance of transcripts. The method is based on the kinetics of DNA hybridization and the unique features of duplex-specific nuclease from the Kamchatka crab. The efficiency of the method was confirmed in a number of model experiments.

Keywords

bacterial transcriptome duplex-specific nuclease depletion of ribosomal RNA subtractive hybridization 

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Copyright information

© Pleiades Publishing, Ltd. 2011

Authors and Affiliations

  • E. A. Bogdanova
    • 1
  • I. A. Shagina
    • 2
  • Yu. G. Yanushevich
    • 2
  • L. L. Vagner
    • 1
    • 2
  • S. A. Lukyanov
    • 1
  • D. A. Shagin
    • 1
    • 2
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.ZAO EvrogenMoscowRussia

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