PCR to genotype RaDR mouse from ear punch
Jennifer Kay, PhD. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA USA
jekay@mit.eduOTHER AUTHORS:
Michelle Sukup Jackson, PhD. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA USA
Joshua Corrigan, MS. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA USA
PROTOCOL TYPE:
Protocol Document
SYNOPSIS:
These protocols describe the process to genotype for the RaDR transgene in mouse ear punches. RaDR mice were developed in the Engelward lab at MIT, and they allow detection of sequence rearrangement mutations arising from aberrant homologous recombination in situ. Please see the paper below for more information on RaDR.
The first protocol describes a quick tissue lysate procedure and the PCR recipe utilizes the commercial Platinum Supermix.
The second protocol in the document describes a tissue digestion procedure that yields purified DNA, and it provides a PCR recipe using individual ingredients that may be more cost-effective.
Sukup-Jackson MR, Kiraly O, Kay JE, Na L, Rowland EA, Winther KE, et al. (2014) Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo. PLoS Genet 10(6): e1004299. doi:10.1371/journal.pgen.1004299