Equalizing cDNA subtraction based on selective suppression of polymerase chain reaction: cloning of Jurkat cell transcripts induced by phytohemaglutinin and phorbol 12-myristate 13-acetate.

Journal Article (Journal Article)

The major drawback of subtractive cDNA libraries is that the original disproportion in concentrations of different types of transcripts is preserved. This usually makes the isolation of specific rare transcripts extremely difficult. To overcome this difficulty, we propose a strategy that introduces the equalization of concentrations (normalization) of specific transcripts during the subtractive process. This makes possible obtaining both rare and highly abundant transcripts in the resulting subtracted library. This technique has been applied for isolation of transcripts activated upon induction of Jurkat cells by phytohemaglutinin and phorbol 12-myristate 13-acetate. Six novel up-regulated sequences belonging to a low-abundance class of transcripts have been obtained.

Full Text

Duke Authors

Cited Authors

  • Gurskaya, NG; Diatchenko, L; Chenchik, A; Siebert, PD; Khaspekov, GL; Lukyanov, KA; Vagner, LL; Ermolaeva, OD; Lukyanov, SA; Sverdlov, ED

Published Date

  • August 15, 1996

Published In

Volume / Issue

  • 240 / 1

Start / End Page

  • 90 - 97

PubMed ID

  • 8811883

International Standard Serial Number (ISSN)

  • 0003-2697

Digital Object Identifier (DOI)

  • 10.1006/abio.1996.0334

Language

  • eng

Conference Location

  • United States