Multiple isoform recovery (MIR)-PCR: a simple method for the isolation of related mRNA isoforms.

Abstract:

:We present a rapid and efficient method for the detection of related transcripts with different expression levels. This approach combines the rapid amplification of cDNA ends (RACE) method with a cDNA subtractive technique. The strategy is based on successive subtractions of prevalent isoforms resulting in enrichment of less expressed transcripts. For each subtraction, a biotinylated primer specific for the prevalent isoform is hybridized on the total cDNA and the hybrid is retained on a streptavidin affinity column. The unbound cDNA serves as a template for subsequent isoform identification. To illustrate its application we describe the isolation of three new actin cDNA isoforms in the freshwater planarian Dugesia (S) polychroa.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Fagotti A,Gabbiani G,Pascolini R,Neuville P

doi

10.1093/nar/26.8.2031

subject

Has Abstract

pub_date

1998-04-15 00:00:00

pages

2031-3

issue

8

eissn

0305-1048

issn

1362-4962

pii

gkb341

journal_volume

26

pub_type

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