Usage Restrictions
In publishing, an acknowledgment to the DEPOSITOR is requested.
Origin
The rats were originally generated by the University of Michigan Transgenic Animal Model Core, University of Michigan. They use a Crl:CD(SD) sub strain of Sprague-Dawley rats, obtained
from Charles River Laboratory (Wilmington, MA, USA). The F1 rats have been bred in the UK with Hsd:Sprague Dawley (Harlan laboratories, UK), which are direct descendants of the original 1925 SD-company colony (Madison, Wisconsin, USA).
Strain characteristics
This rat carries a transgene consisting of the Dsp promoter followed by a 'STOP’ cassette flanked by FRT sites. The stop cassette consists of an mcherry gene encoding for a red fluorescent protein and a kanamycin resistant gene and three SV40polyA adenylation signals. The STOP cassette is followed by a gene encoding for the human diphtheria toxin receptor. Red fluorescence can be detected from the skin of the animals but not the brain.
Breeding Conditions
Both males and fertiles are fertile and generate good litter sizes of 10-15 pups.
Genotyping
The transgene can be detected using the following primers and PCR protocol:
DSP+DTR_gno2_F cca agc tga agg tga cca ag
DSP+DTR_gno2_R tgg tgt agt cct cgt tgt gg
Reagent Volume (ul)
Template DNA (10ng/ul) 2
Forward primer (10mMol) 1
Reverse Primer (10mMol) 1
Buffer (10x) 2
dNTP (8mMol) 2
Mg2+ (25mMol) 1.6
Taq Polymerase (Hot Star, Qiagen) 0.1
ddH20 10.3
Total Volume 20
Step Temp (°C) Time DNA activity
1 98 15 minutes Activation of polymerase
2 95 30s Denaturation
3 64 30s Annealing
4 -0.5 °C per cycle
5 72 1 min Extension
6 Go to step 3, 13 times
7 57 30s Annealing
9 72 1 min Extension
10 Got to step 7, 30 times
11 72 10 min final extension
12 4 forever