Protocol 20627: QPCR Assay - Generic Cre Quantitative PCR

STOCK Tg(mI56i-cre,EGFP)1Kc/J

Stock No: 023724

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-Synb^em1(cre)Mdf/J
004302	129S1/Sv-Hprt1^tm1(CAG-cre)Mnn/J
032051	B6(129S4)-Kit^tm1.1(cre)Jmol/J
037511	B6(129S4)-Pou2f3^tm1.1(cre/ERT2)Imt/J
024976	B6(129S4)-Tspo^tm1.1Maf/J
038514	B6(129S6)-Tg(Slc6a5-cre/ERT2)#Uze/J
017562	B6(Cg)-Cd8a^tm1.1(cre)Koni/J
026512	B6(Cg)-Tg(TPO-cre/ERT2)1139Tyj/J
037052	B6(FVB)-Tg(Sprr2f-cre)2Dcas/J
008320	B6.129(Cg)-Lepr^tm2(cre)Rck/J
033964	B6.129(Cg)-Ptf1a^tm2(cre/ESR1)Cvw Kras^tm4Tyj Pten^tm1Hwu/J
036564	B6.129(Cg)-Scn10a^tm2(cre)Jwo/TjpJ
008463	B6.129-Gt(ROSA)26Sor^tm1(cre/ERT2)Tyj/J
025567	B6.129-Olig2^tm1.1(cre)Wdr/J
004146	B6.129-Tg(Pcp2-cre)2Mpin/J
027038	B6.129S(SJL)-Cyp19a1^tm2.1(cre)Shah/J
032621	B6.129S-Ly6a^tm1.1(cre)Jmol/J
006054	B6.C-Tg(CMV-cre)1Cgn/J
008601	B6.Cg-7630403G23Rik^Tg(Th-cre)1Tmd/J
006230	B6.Cg-Cebpa^tm1Dgt Tg(Mx1-cre)1Cgn/J
036450	B6.Cg-Foxl1^tm2(cre/ERT2)Khk/Mmjax
032923	B6.Cg-Kit^tm1(cre)Htng/J
035169	B6.Cg-Kras^tm4Tyj Apc^tm1Tno Tg(CDX2-cre/ERT2)752Erf/MaraJ
028536	B6.Cg-Ndnf^tm1.1(folA/cre)Hze/J
030544	B6.Cg-Npy1r^tm1.1(cre/GFP)Rpa/J
031485	B6.Cg-Pou4f2^tm1(cre)Bnt/SjJ
003574	B6.Cg-Speer6-ps1^Tg(Alb-cre)21Mgn/J
022865	B6.Cg-Trib2^tm1.1(cre/ERT2)Hze/J
012631	B6.Cg-Tyr^c-2J Sharpin^tm1.1Sun/J
032490	B6.Cg-Wnt4^tm3(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 Braf^tm1Mmcm Pten^tm1Hwu/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-Six2^tm3(EGFP/cre/ERT2)Amc/J
023525	B6;129S-Snap25^tm2.1(cre)Hze/J
036300	B6;129S4-Cux1^tm1(cre/ERT2)Zjh/J
017495	B6;129S7-Crim1^tm1(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-Tmem163^Tg(ACTB-cre)2Mrt/CjDswJ
003465	BALB/c-Tg(CMV-cre)1Cgn/J
004126	C.Cg-Cd19^tm1(cre)Cgn Igh^b/J
035489	C57BL/6-Elmo1^tm1Mki/J
038053	C57BL/6-Oprm1^tm2.1(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-Gne^tm1Dar/J
032080	C57BL/6J-Tg(Cd8a*-cre)B8Asin/J
027789	C57BL/6N-Fgd5^tm3(cre/ERT2)Djr/J
023453	C57BL/6N-Lhx8^tm1(cre)Grsr/2GrsrJ
012929	FVB(Cg)-Tg(Dhh-cre)1Mejr/J
006954	FVB-Tg(Ddx4-cre)1Dcas/J
005025	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd 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-Prkdc^scid Il2rg^tm1Wjl Tg(CAG-cre/Esr1*)5Amc/J
034795	NOD.Cg-Tg(CAG-cre/Esr1*)5Amc/J
035185	STOCK Adgrl1^tm2c(EUCOMM)Hmgu/SudJ
036942	STOCK Ager^tm2.1(cre/ERT2)Blh/2J
012899	STOCK Agrp^tm1(cre)Lowl/J
037028	STOCK Calcr^tm1.1(cre)Mgmj/J
008783	STOCK Grm7^Tg(SMN2)89Ahmb Smn1^tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J
035978	STOCK Hoxb8^tm2.1(cre)Mrc/J
032429	STOCK Kras^tm4Tyj Trp53^tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
026925	STOCK Krt19^tm1(cre/ERT)Ggu/J
018418	STOCK Lrig1^tm1.1(cre/ERT2)Rjc/J
004192	STOCK Mttp^tm2Sgy Ldlr^tm1Her Apob^tm2Sgy Tg(Mx1-cre)1Cgn/J
037139	STOCK Myl1^tm1(cre)Sjb Atg7^tm1Tchi Gaa^tm1Rabn/PuroMmjax
035303	STOCK Obscn^tm1Chen/J
035925	STOCK Opn4^tm1(cre)Saha/J
035045	STOCK Oprk1^tm1.1(cre)Sros/J
030543	STOCK Oxtr^tm1(cre/GFP)Rpa/J
020080	STOCK Slc6a3^tm1(cre)Xz/J
036299	STOCK Tbr1^tm1(cre/ERT2)Zjh/J
036906	STOCK Tmem178b^tm1c(KOMP)Wtsi Tmem178^tm1.1Sud/J
010911	STOCK Wt1^tm1(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