Japanese
 NBRP Rat No: 0640 Strain Name: SD-Tg(Dsp-mCherry,-DTR)Jog Commmon Name: DSP-STOP-hDTR
 Principal Investigator  Jonathan flint
 Organization   University of Oxford  The Wellcome Trust Centre for human Genetics
 Address  Roosevelt Drive

OX3 7BN  Oxford

  UK
 Telephone  +44 (0)1865 287772  Fax:  +44 (0)1865 287501   jf@well.ox.ac.uk
 Inbred Generations   More than 5 
   
 Coat Color
 Deposition Status
 
 White
  Embryo      Sperm      Live Animals
 Usage Restrictions  In publishing, an acknowledgment to the DEPOSITOR is requested.  
 Genetic Status   Inbred   Segregating   Congenic   Consomic    Recombinant 
  Coisogenic   Spont. Mutant    Transgene   Ind. Mutant    Others 
 Comercial Availability   
 Research Category   Diabetes Obesity    Neurobiology    Ophthalmology    Dentistry    Cardio- Hypertension 
  Oncology   Metabolism   Otorhinology    Immunology    Infectious Disease
  Osteology    Internal Medicine   Dermatology   Reproduction    Development
  Behavior    Hematology    Urology   Pharmacology   Others 
  Control Strains   Reporter gene Strains  
 Gene Dsp
 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
 
 References