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

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, effectivel...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry 2005-03, Vol.31 (2), p.170-177
Main Authors: Zhulidov, P. A., Bogdanova, E. A., Shcheglov, A. S., Shagina, I. A., Wagner, L. L., Khazpekov, G. L., Kozhemyako, V. V., Lukyanov, S. A., Shagin, D. A.
Format: Article
Language:English
Subjects:
Decapoda
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Summary: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.
ISSN:1068-1620
1573-9163
1608-330X
DOI:10.1007/s11171-005-0023-7