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Biochem Biophys Res Commun. 1997 Jan 13;230(2):285-8.

Construction of cDNA libraries from small amounts of total RNA using the suppression PCR effect.

Author information

1
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Moscow.

Abstract

Here we describe a method for preparing high-quality cDNA libraries from total RNA. By this method, double-stranded (ds) cDNA ligated with a specially designed ds adaptor is amplified by PCR using a modified T-primer and another primer corresponding to the outer part of the adaptor. The suppression PCR effect strongly inhibits the amplification of poly(A) RNA, thereby reducing background. This method leads to amplification of high-quality cDNA, facilitating the construction of representative cDNA libraries from as little as 10-100 ng of total RNA.

PMID:
9016767
DOI:
10.1006/bbrc.1996.5948
[Indexed for MEDLINE]

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