Stock Number:013149
Strain Name:B6.129-Dysftm1Kcam/J
Neo Qpcr
Generic Neo Quantitative PCR-QPCR- 1.2
QPCR
4.0
22-December -2011 22-December -2011
Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler
480. 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
13741 6-FAM CAT CGC ATC GAG CGA GCA CGT ACT

Zen probe with Iowa Black

 Probe
13762 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A

Zen probe with Iowa Black

 IC Probe
oIMR1080 - CGG CTG CAT ACG CTT GAT C - Forward
oIMR1081 - CGA CAA GAC CGG CTT CCA T - Reverse
oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward
oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
5 uM 13741 0.75 0.15 uM 0.75
5 uM 13762 0.75 0.15 uM 0.75
2 X PerfeCTa qPCR SuperMix 10.00 0.80 X 10.00
40 uM oIMR1080 0.25 0.40 uM 0.25
40 uM oIMR1081 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
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
002356 C57BL/6J-Tg(pPGKneobpA)3Ems/J
004861 B6EiC3Sn-Ts(16C-tel)1Cje/DnJ
005032 B6 x B6Ei.129(Cg)-Bmp8btm1Blh/EiJ
005557 NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
006021 NOD.Cg-H2-Ab1tm1Gru Tg(CD2-CD4,HLA-DQA1,HLA-DQB1)1Ell/EllJ
006022 NOD.Cg-Rag1tm1Mom H2-Ab1tm1Gru Tg(CD2-CD4,HLA-DQA1,HLA-DQB1)1Ell/EllJ
006023 NOD.Cg-H2-Ab1tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ
006024 NOD.Cg-Rag1tm1Mom H2-Ab1tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ
006378 B6;129-Nrxn2tm3Sud/J
006830 129-Dysftm1Kcam/J
009340 STOCK Eif2ak3tm1Dron/HotaJ
010966 NODCaj.129S2(B6)-H2-Ab1tm1Doi/LwnJ
013149 B6.129-Dysftm1Kcam/J
017637 NOD.Cg-Prkdcscid Il2rgtm1Wjl H2-Ab1tm1Gru Tg(HLA-DRB1)31Dmz/SzJ
019578 B6;129S1-Rnlstm1Gvd/J
350108 PL/J x B6.129S7-Itgb2tm1Bay/J
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.