|Method of Cryopreservation||Sperm|
|Trial IVF % Fertilization||74.63%|
|Mutation #1: Targeted Mutagenesis|
|Zygosity at cryopreservation||Heterozygote|
|PCR Genotyping Protocol||Sst.rtTA_PCR_genotyoping_protocol.docx|
|Strain Type||Congenic Strain|
|Chimera/Founder Genetic Background||129S6/SvEvTac|
|Cryopreservation Strain Background (VCMR)||C57BL/6J|
|Viability and Fertility Data||
Strain background: 96.8% C57Bl6/J at cryopreservation.
Expected to be homozygous viable and fertile.
Heterozygous viable and fertile.
The targeting vector contains 7.3 Kb 5’ and a 3.6 kb 3’ homology arms. Lox66 and Lox2272 sites were inserted around a 500 bp region of the of Sst promoter and exons I and II of the somatostatin gene. rTTA coding sequences with a beta-globin polyA site replaced exons I and II of the Sst. The vector also contains FRT-flanked puTK-EM7 Neo selection double selection cassette. PU(delta)TK was used for positive selection for targeting events with puromycin and negative selection for RMCE events with ganciclovir. EM7-Neo was used for positive selection in bacteria during BAC recombineering process.