Accumulated background variation among H2 mutant congenic strains : elimination through PCR-based genotyping of F2 segregants

摘要: Abstract Many commercially and privately available congenic strains of laboratory animals were founded decades ago are likely to differ from one another by dozens fixed mutational differences at background loci. This problem is often ignored despite growing evidence that such variation exists. Eliminating this confounding can be largely accomplished crossing produce F2 segregants homozygous (or heterozygous) for relevant genes. Discriminating homozygotes difficult when strain minor, as mutant mouse differing single major histocompatibility loci (H2 congenics). Here, we describe a two-step polymerase chain reaction (PCR) method utilizing heteroduplex analysis sequence specific primers (SSP-PCR) efficiently discriminates the progeny two H2 mice crosses (bm1×B6 bm1×bm3). A third cross cannot resolved heteroduplexing, but discriminated (albeit less efficiently) with SSP-PCR alone. sensitive application extended any strains.