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

IPC = 74 bp

Tg = 102 bp

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
038200 129S;FVB-Synbem1(cre)Mdf/J
004302 129S1/Sv-Hprt1tm1(CAG-cre)Mnn/J
032051 B6(129S4)-Kittm1.1(cre)Jmol/J
037511 B6(129S4)-Pou2f3tm1.1(cre/ERT2)Imt/J
024976 B6(129S4)-Tspotm1.1Maf/J
038514 B6(129S6)-Tg(Slc6a5-cre/ERT2)#Uze/J
017562 B6(Cg)-Cd8atm1.1(cre)Koni/J
026512 B6(Cg)-Tg(TPO-cre/ERT2)1139Tyj/J
037052 B6(FVB)-Tg(Sprr2f-cre)2Dcas/J
008320 B6.129(Cg)-Leprtm2(cre)Rck/J
033964 B6.129(Cg)-Ptf1atm2(cre/ESR1)Cvw Krastm4Tyj Ptentm1Hwu/J
036564 B6.129(Cg)-Scn10atm2(cre)Jwo/TjpJ
008463 B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J
025567 B6.129-Olig2tm1.1(cre)Wdr/J
004146 B6.129-Tg(Pcp2-cre)2Mpin/J
027038 B6.129S(SJL)-Cyp19a1tm2.1(cre)Shah/J
032621 B6.129S-Ly6atm1.1(cre)Jmol/J
008601 B6.Cg-7630403G23RikTg(Th-cre)1Tmd/J
006230 B6.Cg-Cebpatm1Dgt Tg(Mx1-cre)1Cgn/J
036450 B6.Cg-Foxl1tm2(cre/ERT2)Khk/Mmjax
032923 B6.Cg-Kittm1(cre)Htng/J
035169 B6.Cg-Krastm4Tyj Apctm1Tno Tg(CDX2-cre/ERT2)752Erf/MaraJ
028536 B6.Cg-Ndnftm1.1(folA/cre)Hze/J
030544 B6.Cg-Npy1rtm1.1(cre/GFP)Rpa/J
031485 B6.Cg-Pou4f2tm1(cre)Bnt/SjJ
038835 B6.Cg-Sdk1em1(cre/GFP)Jrs/KrishJ
003574 B6.Cg-Speer6-ps1Tg(Alb-cre)21Mgn/J
022865 B6.Cg-Trib2tm1.1(cre/ERT2)Hze/J
012631 B6.Cg-Tyrc-2J Sharpintm1.1Sun/J
032490 B6.Cg-Wnt4tm3(EGFP/cre)Amc/J
006149 B6.Cg-Tg(ACTA1-cre)79Jme/J
030215 B6.Cg-Tg(Amigo2-cre)1Sieg/J
022357 B6.Cg-Tg(Arc-cre/ERT2)MRhn/CdnyJ
005359 B6.Cg-Tg(Camk2a-cre)T29-1Stl/J
006368 B6.Cg-Tg(Cr2-cre)3Cgn/J
029594 B6.Cg-Tg(Dmp1-cre/ERT2)D77Pdp/J
006663 B6.Cg-Tg(Eno2-cre)39Jme/J
035668 B6.Cg-Tg(Gata1-cre)1Sho/MdfJ
012886 B6.Cg-Tg(Gfap-cre)73.12Mvs/J
024098 B6.Cg-Tg(Gfap-cre)77.6Mvs/2J
003573 B6.Cg-Tg(Ins2-cre)25Mgn/J
003771 B6.Cg-Tg(Nes-cre)1Kln/J
008205 B6.Cg-Tg(NPHS2-cre)295Lbh/J
008827 B6.Cg-Tg(Prdm1-cre)1Masu/J
005584 B6.Cg-Tg(Prrx1-cre)1Cjt/J
021614 B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J
003966 B6.Cg-Tg(Syn1-cre)671Jxm/J
004128 B6.Cg-Tg(Tek-cre)12Flv/J
008863 B6.Cg-Tg(Tek-cre)1Ywa/J
029788 B6.Cg-Tg(Tyr-cre)1Lru/J
013590 B6.Cg-Tg(Tyr-cre/ERT2)13Bos Braftm1Mmcm Ptentm1Hwu/BosJ
035670 B6.Cg-Tg(VAV1-cre)1Graf/MdfJ
020282 B6.Cg-Tg(Vil1-cre/ERT2)23Syr/J
008735 B6.Cg-Tg(Wap-cre)11738Mam/JKnwJ
006137 B6.FVB-Tg(Cdh5-cre)7Mlia/J
018980 B6.FVB-Tg(Ddx4-cre)1Dcas/KnwJ
003724 B6.FVB-Tg(EIIa-cre)C5379Lmgd/J
011038 B6.FVB-Tg(Myh6-cre)2182Mds/J
010714 B6.FVB-Tg(Pomc-cre)1Lowl/J
009600 B6;129-Six2tm3(EGFP/cre/ERT2)Amc/J
023525 B6;129S-Snap25tm2.1(cre)Hze/J
036300 B6;129S4-Cux1tm1(cre/ERT2)Zjh/J
017495 B6;129S7-Crim1tm1(GFP/cre/ERT2)Pzg/J
009613 B6;C3-Tg(Scnn1a-cre)3Aibs/J
009103 B6;C3-Tg(Wfs1-cre/ERT2)3Aibs/J
027784 B6;CBA-Tg(Emx1-cre/ERT2)1Kess/SshiJ
025809 B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J
015855 B6;DBA-Tg(Upk3a-GFP/cre/ERT2)26Amc/J
023748 B6;FVB-Tg(Aldh1l1-cre)JD1884Htz/J
004426 B6;SJL-Tg(Cga-cre)3Sac/J
003554 B6;SJL-Tg(Col2a1-cre)1Bhr/J
003552 B6129-Tg(Wap-cre)11738Mam/J
018965 B6N.Cg-Tg(Fabp4-cre)1Rev/J
018964 B6N.Cg-Tg(KRT14-cre)1Amc/J
019099 B6N.FVB-Tmem163Tg(ACTB-cre)2Mrt/CjDswJ
003465 BALB/c-Tg(CMV-cre)1Cgn/J
004126 C.Cg-Cd19tm1(cre)Cgn Ighb/J
035489 C57BL/6-Elmo1tm1Mki/J
038053 C57BL/6-Oprm1tm2.1(EGFP/cre)Ics/KffJ
017557 C57BL/6-Tg(BEST1-cre)1Jdun/J
016097 C57BL/6-Tg(Car1-cre)5Flt/J
016261 C57BL/6-Tg(Nes-cre/ERT2)KEisc/J
003651 C57BL/6-Tg(Zp3-cre)93Knw/J
036429 C57BL/6J-Gnetm1Dar/J
032080 C57BL/6J-Tg(Cd8a*-cre)B8Asin/J
027789 C57BL/6N-Fgd5tm3(cre/ERT2)Djr/J
023453 C57BL/6N-Lhx8tm1(cre)Grsr/2GrsrJ
012929 FVB(Cg)-Tg(Dhh-cre)1Mejr/J
006954 FVB-Tg(Ddx4-cre)1Dcas/J
005025 FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb/J
018394 FVB.Cg-Tg(KRT5-cre/ERT2)2Ipc/JeldJ
003314 FVB/N-Tg(EIIa-cre)C5379Lmgd/J
034859 NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-cre/Esr1*)5Amc/J
034795 NOD.Cg-Tg(CAG-cre/Esr1*)5Amc/J
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
036942 STOCK Agertm2.1(cre/ERT2)Blh/2J
012899 STOCK Agrptm1(cre)Lowl/J
037028 STOCK Calcrtm1.1(cre)Mgmj/J
008783 STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J
035978 STOCK Hoxb8tm2.1(cre)Mrc/J
032429 STOCK Krastm4Tyj Trp53tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
026925 STOCK Krt19tm1(cre/ERT)Ggu/J
018418 STOCK Lrig1tm1.1(cre/ERT2)Rjc/J
004192 STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J
037139 STOCK Myl1tm1(cre)Sjb Atg7tm1Tchi Gaatm1Rabn/PuroMmjax
035303 STOCK Obscntm1Chen/J
035925 STOCK Opn4tm1(cre)Saha/J
035045 STOCK Oprk1tm1.1(cre)Sros/J
030543 STOCK Oxtrtm1(cre/GFP)Rpa/J
020080 STOCK Slc6a3tm1(cre)Xz/J
036299 STOCK Tbr1tm1(cre/ERT2)Zjh/J
036906 STOCK Tmem178btm1c(KOMP)Wtsi Tmem178tm1.1Sud/J
010911 STOCK Wt1tm1(EGFP/cre)Wtp/J
035566 STOCK Tg(CAG-mRuby2*,-mClover*)1Srgt/J
005105 STOCK Tg(Chx10-EGFP/cre,-ALPP)2Clc/J
012841 STOCK Tg(Ggt1-cre)M3Egn/J
027681 STOCK Tg(Hnf1b-cre/ERT2)1Jfer/J
023426 STOCK Tg(Kiss1-cre)J2-4Cfe/J
004782 STOCK Tg(KRT14-cre)1Amc/J
005107 STOCK Tg(KRT14-cre/ERT)20Efu/J
023724 STOCK Tg(mI56i-cre,EGFP)1Kc/J
003553 STOCK Tg(MMTV-cre)4Mam/J
028364 STOCK Tg(Neurod1-cre)1Able/J
032646 STOCK Tg(Spo11-cre)1Rsw/PecoJ
025520 STOCK Tg(T-cre/ERT2)1Lwd/J
004746 STOCK Tg(Tagln-cre)1Her/J
016584 STOCK Tg(Tph2-icre/ERT2)6Gloss/J
008851 STOCK Tg(Wnt1-cre/ERT)1Alj/J
128 strains use this protocol