Transgene Assay Form

This form must be completed prior to initiation of animal work.

Investigator: _______________________  Transgene or Locus Name: ____________________

Assay Method:       ______ PCR                                          ______ Southern

Instructions

            PCR Assay - The PCR assay must show the results of a positive control (typically the transgene vector alone), a negative control (typically water) and a transgene-genomic DNA mixture. The transgene-genomic DNA mixture is made by combining the transgene plasmid DNA with mouse genomic DNA to final concentrations of 300 fentograms/microliter (for a 10 kb plasmid) and 100 nanograms/microliter, respectively. Use 1 microliter of the mixture per PCR reaction. A picture of the gel indicating the lane assignments must be attached below.

            Southern Assay - The Southern assay must show the results of a blot in which 30pg of the vector is mixed with 10 micrograms of mouse genomic DNA (available from the facility) and cut simultaneously with restriction endonuclease(s).  A picture of the results  must be attached below.