Russian Journal of Bioorganic Chemistry

, Volume 31, Issue 2, pp 170–177

A method for the preparation of normalized cDNA libraries enriched with full-length sequences

  • P. A. Zhulidov
  • E. A. Bogdanova
  • A. S. Shcheglov
  • I. A. Shagina
  • L. L. Wagner
  • G. L. Khazpekov
  • V. V. Kozhemyako
  • S. A. Lukyanov
  • D. A. Shagin
Article

Abstract

We developed a new method for the preparation of normalized cDNA libraries enriched with full-length sequences. It is based on the properties of the recently characterized duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. The duplex-specific nuclease is thermostable, effectively cleaves double-stranded DNA, and is inactive toward single-stranded DNA (Shagin et al., Genome Res., 2002, vol. 12, pp. 1935–1942). Our method enables the normalization of cDNA samples enriched with full-length sequences without use of laborious and ineffective stages of physical separation. The efficiency of the method was demonstrated in model experiments using cDNA samples from several human tissues.

Key words

cDNA normalization duplex-specific nuclease normalized cDNA libraries 

Abbreviations

ds cDNA

double-stranded cDNA

DSN

duplex-specific nuclease

ITR

inverted terminal repeat

PCR

polymerase chain reaction

SMART

switching mechanism at the 5′-end of the RNA transcript

ss cDNA

single-stranded cDNA

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Copyright information

© MAIK “Nauka/Interperiodica” 2005

Authors and Affiliations

  • P. A. Zhulidov
    • 1
  • E. A. Bogdanova
    • 1
  • A. S. Shcheglov
    • 1
  • I. A. Shagina
    • 2
  • L. L. Wagner
    • 1
    • 2
  • G. L. Khazpekov
    • 3
  • V. V. Kozhemyako
    • 4
  • S. A. Lukyanov
    • 1
  • D. A. Shagin
    • 1
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.ZAO EvrogenMoscowRussia
  3. 3.Cardiological Research CenterRussian Academy of Medical SciencesMoscowRussia
  4. 4.Pacific Institute of Bioorganic Chemistry, Far East DivisionRussian Academy of SciencesVladivostokRussia

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