Strain distribution patterns for 50 SSLP markers in the murine AKXL recombinant inbred set

Abstract

The ability of a recombinant inbred (RI) strain set to detect linkage is dependent on the number of strains in the set and the number of genetic markers typed for the set (Neumann 1990; Taylor 1978). Of the two options, it is easier, and more practical, to add markers to a set than it is to add new strains. Using simple sequence length polymorphism (SSLP) markers developed and mapped by Dietrich (Dietrich et al. 1992a,b,c), we obtained the strain distribution patterns (SDPs) of 50 additional loci for the AKXL RI set derived from the mouse inbred strains AKR/J and C57L/J. These loci were chosen because of their relevance to another project, hence they are concentrated in five chromosomal regions. This brings the number of loci typed in this RI set to over 300. PCR primers for the SSLP markers were purchased from Research Genetics (Huntsville, Ala.). Genomic DNA was purchased from The Jackson Laboratory DNA Resource and diluted to a concentration of 100 ng/gl with TE (10 mM Tris-HC1 (pH 8.0), 1 mM EDTA) before use. PCR was carried out in a 14-gl volume. The final concentration for each reaction was: 10 mM Tris-HC1 (pH 8.3), 50 mM KC1, 1.9 mM MgC12, 200 gM dNTP, 0.025 U/~tl AmpliTaq DNA polymerase (Perkin Elmer Cetus, Norwalk, Conn.), 0.132 gM of each primer, and 7.5 ng/gl DNA template. Amplification was carried out as described by Eicher and Shown (1993). PCR products were then separated in a 4% Nusieve-agarose (1:1) gel in 1 • TBE for 6 h at 70V (2.4 V/cm). TBE and gel loading buffer were prepared and