Extended Data Figure 2. Generation of Mycl1^gfp/gfp mice by homologous recombination.

a, The genomic context for Mycl1, the targeting vector, and targeted allele are shown. NotI restriction digest of the WT locus generates a 12.5 kb fragment detectable by either 5' or 3' probes. NcoI restriction digest of the targeted locus generates a 3.3 kb fragment detected by the 5' probe, a 2.3 kb fragment containing most of the GFP-Neo cassette, and a 9.3 kb fragment detected by the 3' probe. b, c, Southern blot analysis of NcoI-digested total DNA using the 5' probe (b) and 3' probe (c). Shown are results obtained from genomic DNA of WT (Mycl1^+/+) and heterozygous (Mycl1^+/gfp) mice. d, Genotyping PCR using genomic DNA of WT (Mycl1^+/+), heterozygous (Mycl1^+/gfp), and homozygous (Mycl1^gfp/gfp) mice. The WT allele yields a 337 bp product and the targeted allele yields a 525 bp product.