Stock Number:006604
Strain Name:NOD/ShiLtDvs-Tg(HLA-A/H2-D/B2M)1Dvs/J
Tg(HLA-A/H2-D/B2M)1Dvs
Tg(HLA-A/H2-D)
QPCR
3.0
29-October -2010 21-July -2011
Notes:

 

Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate
instrument specific Fluorophore/Quencher combination. The transgene genotype is
determined by comparing ΔCt values of each unknown sample against known
homozygous and hemizygous controls, using appropriate endogenous references.

 
Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
11287 - GGA GAC ACG GAA AGT GAA GG - Transgene Forward
11288 - GGC CTC GCT CTG GTT GTA G - Transgene Reverse
11289 6-FAM CCG AGT GGA CCT GGG GAC CC Black Hole Quencher 1 Tg Probe
oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward
oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse
TmoIMR0105 Cy5 CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 2 IC Probe
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
2 X PerfeCTa qPCR SuperMix 10.00 0.80 X 10.00
40 uM oIMR4310 0.25 0.40 uM 0.25
40 uM oIMR4311 0.25 0.40 uM 0.25
40 uM oIMR1544 0.25 0.40 uM 0.25
40 uM oIMR3580 0.25 0.40 uM 0.25
5 uM TmoIMR0070 0.75 0.15 uM 0.75
5 uM TmoIMR0107 0.75 0.15 uM 0.75
ddH2O 2.50 - 2.50
DNA 10.00 - 10.00
Cycling 
Step # Temp °C Time Note
1 50 2 min -
2 95 10 min -
3 95 15 sec -
4 60 1 min repeat steps 3-4 for 40 cycles
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
003475 C57BL/6-Tg(HLA-A2.1)1Enge/J
004191 B6.Cg-Tg(HLA-A/H2-D)2Enge/J
006604 NOD/ShiLtDvs-Tg(HLA-A/H2-D/B2M)1Dvs/J
006605 NOD.Cg-Prkdcscid Emv30b Tg(HLA-A/H2-D/B2M)1Dvs/DvsJ
006611 NOD.129P2(B6)-B2mtm1Unc Tg(HLA-A/H2-D/B2M)1Dvs/DvsJ
009617 NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(HLA-A2.1)1Enge/SzJ
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.