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

Konstantin Lukyanov, Luda Diatchenko, Alex Chenchik, Amulia Nanisetti, Paul Siebert, Natalia Usman, Mikhail Matz, Sergey Lukyanov

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)285-288
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume230
Issue number2
DOIs
Publication statusPublished - 13 Jan 1997
Externally publishedYes

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