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

K Lukyanov; L Diatchenko; A Chenchik; A Nanisetti; P Siebert; N Usman; M Matz; S Lukyanov

doi:10.1006/bbrc.1996.5948

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

Biochem Biophys Res Commun. 1997 Jan 13;230(2):285-8. doi: 10.1006/bbrc.1996.5948.

Authors

K Lukyanov¹, L Diatchenko, A Chenchik, A Nanisetti, P Siebert, N Usman, M Matz, S Lukyanov

Affiliation

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

PMID: 9016767
DOI: 10.1006/bbrc.1996.5948

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.

MeSH terms

DNA, Complementary* / isolation & purification
Electrophoresis, Agar Gel
Gene Library*
Humans
Muscle, Skeletal / metabolism*
Polymerase Chain Reaction / methods*
RNA*

Substances

DNA, Complementary
RNA