Humanized Gene Knock-in Strategy
Figure 1. Targeting strategy for Trem2 gene CKO mouse model by ES cell targeting method.
Genotyping Strategy
Figure 2. Genotyping strategy for heterozygous F1 mice. Genotyping PCR was done using primer sets for the 5’ arm (regions 1 and 2) and 3’ arm (regions 3 and 4). The 5’ arm PCR yielded a 6.4 kb amplified fragment in the positive F1 mice and a 10.9 kb fragment would be amplified. The 3’ arm PCR yielded a 7.1 kb fragment in F1 positive mice and a 11.1 kb fragment in negative F1 mice.
PCR Gel Electrophoresis to Genotype Recombinant F1 Mice
Figure 3. Agarose gel electrophoresis of PCR results. 5’ recombinant fragment (5'arm ), 3’ recombinant fragment (3'arm ). Six positive F1 mice were identified and confirmed by Sanger sequencing (data not shown). WT wild type control; M: 1kb DNA ladder.