Fig. 3.

Germ line transmission and establishment of a homozygous Cr4 insertion mutation. (A) A sequencing reaction chromatogram from a founder mouse showing a correct donor-oligo insertion at the Cr4 target site. (B) The sequence confirmed mutant was bred with a wild type mouse and germ line transmission was established. Genotyping of F2 generation litter with flanking primers PCR (left) and with Internal + External primers PCR (right). The expected product sizes and the genotypes are given below the gel images. M: 100 base pair Marker. (C) Nucleotide sequence of the locus containing the correct insertion sequence. The locations of primers are shown with the corresponding color coded arrows. Cr4 sequence is underlined and PAM sequence is shown in uppercase.