PCR Genotyping

Genotyping of mutant strains is offered to laboratories using our Husbandry Service.  As an extension of the breeding services, we take tissue biopsy and transfer to our PCR lab for screening.  We use a Universal Polymerase Chain Reaction Assay to optimize the PCR with the same cycling conditions for all samples. For breeding animals, assay results are normally obtained by two weeks of age.  Primers for the Universal Polymerase Chain Reaction Assay must be 30 nucleotides in length, have 40-60% GC content, have non-homologous 3’ ends, contain no repetitive sequences, and produce an amplimer of 100-500 base pairs in length. We request that labs design these long primers and test them in your lab before handing them off for our genotyping service. Please let us know if you have questions on how to design these oligos.

For samples taken in your lab that need testing, please see the submission forms to submit the genotyping request form.  You should include a hard copy of the form with your tissue samples. Tissue should be submitted in 0.2uL PCR strip tubes. We use VWR tubes, cat. #53509-306. For each litter, please clearly label the first tube with a unique identifier, such as AA, which denotes the litter ID. The rest of the tubes should be labeled with the sample numbers (2-6 etc.) on the side of the tube, not the cap, with permanent marker. Samples should be1-3mm in size. Please be sure that you push the tissue sample to the bottom of each tube for the digestion process. Place your strip tubes in an empty pipet box so that the samples remain upright, and tails are not dislodged to the side or top of the tube.