5/27/2023 0 Comments Subler digitalThaw one straw for test of motility or fertilization.Collect and freeze sperm (from 2 males -> ~20 straws).Reimplant 2-cell embryos into foster females.Superovulate (Hyperova) females and collect unfertilized eggs.Transfer rederived mice to investigator. Collect fertilized eggs, transfer to MMRB, and reimplant into foster females.Expand colony if needed purchase/transfer female breeders if needed.Send to Transnetyx (tailed by Investigator).Send to Transnetyx (tailed by Shared Resource).Transfer experimental animals as produced to investigator.Perform all mating, weaning, tail tissue collection, genotyping (usually by Transnetyx), record-keeping, and monitoring of colony.Acquire breeders (from JAX or other vendor, imported from another institution and rederived, or generated by TG/KO) and expand colony.Breed potential founders and screen offspring (including sequencing) to confirm germline transmission of desired allele.Develop genotyping strategies and screen potential founder mice sequencing.Microinjection/electroporation of reagents into fertilized eggs of desired strain and reimplant into foster females.Design targeting strategy (overall model design guide RNA selection repair template selection).KO/KI/CKO Mouse Production by CRISPR/Cas9 $7,000 (for production in C57BL/6J or CB6F1 background additional cost for other backgrounds) A minimum of 3 founders usually generated.
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