Genotyping Protocols Database
Stock No: 003573
Protocol 20627: QPCR Assay - Generic Cre Quantitative PCR
Version 9.0

Notes

Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/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.
The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX). To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility. Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.

Expected Results

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
13593 Fluorophore-1 AAA CAT GCT TCA TCG TCG GTC CGG Quencher-1 Tg Probe Cre probe
oIMR1084 GCG GTC TGG CAG TAA AAA CTA TC Transgene Forward A Cre F
oIMR1085 GTG AAA CAG CAT TGC TGT CAC TT Transgene Reverse A Cre R
oIMR1544 CAC GTG GGC TCC AGC ATT Internal Positive Control Forward A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
TmoIMR0105 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 IC Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
oIMR1084 0.40 uM
oIMR1085 0.40 uM
oIMR1544 0.40 uM
oIMR3580 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 -- repeat steps 2-3 for 40 cycles
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

Stock Number Strain Name
003328 129S/Sv-Tg(Prm-cre)58Og/J
003465 BALB/c-Tg(CMV-cre)1Cgn/J
003553 STOCK Tg(MMTV-cre)4Mam/J
003573 B6.Cg-Tg(Ins2-cre)25Mgn/J
003574 B6.Cg-Speer6-ps1Tg(Alb-cre)21Mgn/J
003651 C57BL/6-Tg(Zp3-cre)93Knw/J
003724 B6.FVB-Tg(EIIa-cre)C5379Lmgd/J
003771 B6.Cg-Tg(Nes-cre)1Kln/J
003966 B6.Cg-Tg(Syn1-cre)671Jxm/J
004146 B6.129-Tg(Pcp2-cre)2Mpin/J
004192 STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J
004426 B6;SJL-Tg(Cga-cre)3Sac/J
004746 STOCK Tg(Tagln-cre)1Her/J
004782 STOCK Tg(KRT14-cre)1Amc/J
005105 STOCK Tg(Chx10-EGFP/cre,-ALPP)2Clc/J
005359 B6.Cg-Tg(Camk2a-cre)T29-1Stl/J
006054 B6.C-Tg(CMV-cre)1Cgn/J
006137 B6.FVB-Tg(Cdh5-cre)7Mlia/J
006230 B6.Cg-Cebpatm1Dgt Tg(Mx1-cre)1Cgn/J
006954 FVB-Tg(Ddx4-cre)1Dcas/J
008766 C57BL/6-Tg(Cd8a-cre)1Itan/J
008827 B6.Cg-Tg(Prdm1-cre)1Masu/J
008851 STOCK Tg(Wnt1-cre/ERT)1Alj/J
009613 B6;C3-Tg(Scnn1a-cre)3Aibs/J
012841 STOCK Tg(Ggt1-cre)M3Egn/J
012886 B6.Cg-Tg(Gfap-cre)73.12Mvs/J
012899 STOCK Agrptm1(cre)Lowl/J
016261 C57BL/6-Tg(Nes-cre/ERT2)KEisc/J
017562 B6(Cg)-Cd8atm1.1(cre)Koni/J
018394 FVB.Cg-Tg(KRT5-cre/ERT2)2Ipc/JeldJ
018964 B6N.Cg-Tg(KRT14-cre)1Amc/J
018980 B6.FVB-Tg(Ddx4-cre)1Dcas/KnwJ
019099 B6N.FVB-Tmem163Tg(ACTB-cre)2Mrt/CjDswJ
022357 B6.Cg-Tg(Arc-cre/ERT2)MRhn/CdnyJ
023426 STOCK Tg(Kiss1-cre)J2-4Cfe/J
023453 C57BL/6N-Lhx8tm1(cre,Cerulean)Grsr/2GrsrJ
023748 B6;FVB-Tg(Aldh1l1-cre)JD1884Htz/J
024098 B6.Cg-Tg(Gfap-cre)77.6Mvs/2J
025520 STOCK Tg(T-cre/ERT2)1Lwd/J
026925 STOCK Krt19tm1(cre/ERT)Ggu/J
027038 B6.129S(SJL)-Cyp19a1tm2.1(cre)Shah/J
027784 B6;CBA-Tg(Emx1-cre/ERT2)1Kess/SshiJ
029594 B6.Cg-Tg(Dmp1-cre/ERT2)D77Pdp/J
029788 B6.Cg-Tg(Tyr-cre)1Lru/J
030757 B6.Cg-Ndnftm1.1(cre)Rudy/J
032429 STOCK Krastm4Tyj Trp53tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
032490 B6.129S4-Wnt4tm3(EGFP/cre)Amc/J
032621 B6.129S-Ly6atm1.1(cre)Jmol/J
032646 STOCK Tg(Spo11-cre)1Rsw/PecoJ
49 strains use this protocol