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. 1998 Apr 15;26(8):2031–2033. doi: 10.1093/nar/26.8.2031

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

A Fagotti 1, G Gabbiani 1, R Pascolini 1, P Neuville 1
PMCID: PMC147508  PMID: 9518500

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.

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Selected References

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