A GENETIC-LINKAGE MAP OF THE MOUSE USING AN EXPANDED PRODUCTION SYSTEM OF RESTRICTION LANDMARK GENOMIC SCANNING (RLGS-VER-1.8)

We have developed an expanded system (RLGS Ver.1.8) for producing RLGS patterns that result in a 16-fold increase in the number of gels prod uced and a 10-fold reduction in the total cost per gel. The major modi fications include: 1) performing the blocking and labeling step withou t phenol extraction or ethanol precipitation; 2) minimizing the reacti on volume and the enzyme units in each step; 3) developing a long vert ical agarose disc gel electrophoresis for the Ist-dimension; and 4) de veloping a new apparatus for multiplex vertical 2nd-dimensional electr ophoresis. RLGS Ver.1.8 was used with a new combination of restriction enzymes to identify variation for 209 loci between C57BL/6J and DBA/2 J. Twenty-six BXD RI strains were analyzed and 195/209 loci were genet ically mapped. These loci were mapped in one week of laboratory work b y two people. This system provides an important tool for the genetic a nalysis of new loci in similar genetic resources. (C) 1994 Academic Pr ess, Inc.