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RT-PCR Primer Sets for Human and Mouse Mismatch Repair Genes

Of each PCR reaction, 10 l was run on a 2% agarose, Tris-acetate gel stained with ethidium bromide.

The expected amplification products from human cDNA ranged from 515 to 876 bp. The primer set for hGTBP produced a significant quantity of PCR product as a result of amplification from genomic template, resulting in a 3.5- to 4-kb band. Since this band migrated more slowly than the 515-bp band produced from amplification of cDNA, it is unlikely to cause problems with RT-PCR analysis of cDNA preparations contaminated with genomic DNA. None of the other primer sets resulted in detectable PCR product when amplifying from human genomic DNA template.

figure 2

A similar experiment contrasted results using the mouse RT-PCR primer sets to amplify either mouse liver cDNA or mouse genomic DNA templates (figure 2). The expected mouse cDNA-derived products in this group ranged from 504 to 926 bp. None of the mouse primer sets amplified a visible product from genomic mouse DNA template when reactions were performed at the recommended annealing temperature.

Conclusions

Characterization of mammalian DNA repair enzymes is a rapidly advancing field. To meet the needs of researchers studying these pathways, Stratagene has developed RT-PCR primers sets for the newly identified genes involved in mammalian DNA mismatch repair. These primers sets amplify regions that span intron-exon boundaries, minimizing the effects of amplification from contaminating genomic DNA template. Each primer set includes resource information for the derivation of the primer set.

REFERENCES
  1. Freidberg, E.C., et al. (1995) In DNA Repair and Mutagenesis, pp.367-368. ASM Press, Washington, D.C.

  2. Karran, P.
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