Figure 1. Production of Mcph1-deficienct (Mcph1^tm1a/tm1a) mice.

(A) Schematic of knockout strategy for Mcph1 gene based on knockout-first design. A promoterless cassette including LacZ and neo genes was inserted in the third intron of Mcph1 gene flanked by FRT sites. LoxP sites flank the critical exon (exon4 of Mcph1 gene in knockout-first design). See http://www.knockoutmouse.org/martsearch/project/41705 for more details. (B) Short range PCR for genotyping. Wild type allele produces one band of 366bp. Due to the insertion of the cassette, primers designed for the wild type allele do not have product for the mutant allele using the short range PCR (illustrated as the left panel, schematic illustration is not in scale). The homozygous allele produces only one band of 185 bp. The heterozygotes produce two bands of 185 bp and 366 bp. (C) Quantitative real-time PCR showed largely reduced transcript of Mcph1 in Mcph1^{tm1a /tm1a} (n = 3) mice compared to wild type mice (n = 3) and the residual levels vary in different organs.