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
002609 B6.129P2-Nos2tm1Lau/J
003124 B6.129-Ucp1tm1Kz/J
004225 B6.129X1-Zap70tm1Weis/J
004861 B6EiC3Sn-Ts(16C-tel)1Cje/DnJ
005032 B6 x B6Ei.129(Cg)-Bmp8btm1Blh/EiJ
005557 NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ
006023 NOD.Cg-H2-Ab1tm1Gru Tg(CD4,HLA-DQA1,HLA-DQB1)N8Ell/EllJ
006378 B6;129-Nrxn2tm3Sud/J
006830 129-Dysftm1Kcam/J
007078 CByJ.129S2(B6)-Il6tm1Kopf/J
009340 B6.129S6(Cg)-Eif2ak3tm1Dron/HotaJ
011131 B6.129X1(Cg)-Arrb1tm1Jse/J
013149 B6.129-Dysftm1Kcam/J
017959 C57BL/6-Pmltm1(PML/RARA)Ley/J
017976 B6.129-Ulk1tm1Thsn/J
018311 B6.129S4-Catsper1tm1Clph/J
018437 B6;129-Pax2tm1Gdr/J
019578 B6;129S1-Rnlstm1Gvd/J
021209 B6;129S1-Kat8tm1Thl/J
022350 MRL.Cg-Nos2tm1Lau Faslpr/J
022760 MRL.Cg-Nos3tm1Unc Faslpr/J
023496 B6.129P3-Rasgrp3tm1Jstn/TbwnJ
024744 B6N.Cg-Idstm1Muen/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.