Methods / RNA isolation and purification
Multiple isoform recovery (MIR)-PCR: a simple method for the isolation of related mRNA isoforms.
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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.
Nucleic Acids Research.1998:26(8):2031
Last update 30-Nov-1999, Rating n/a of 0 votes.
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