Protocol 22392: Standard PCR Assay - Generic Cre

STOCK Kras^tm4Tyj Trp53^tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J

Stock No: 032429

Version 1.3

Notes

This assay will NOT distinguish hemizygous from homozygous transgenic animals.

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

Transgene = ~100 bp
Internal positive control = 324 bp

Separated by gel electrophoresis on a 3.0% agarose gel.

JAX Protocol

Protocol Primers

Primer	5' Label	Sequence 5' → 3'	3' Label	Primer Type	Reaction	Note
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
oIMR7338		CTA GGC CAC AGA ATT GAA AGA TCT		Internal Positive Control Forward	A
oIMR7339		GTA GGT GGA AAT TCT AGC ATC ATC C		Internal Positive Control Reverse	A

Reaction A

Component	Final Concentration
ddH2O
Kapa 2G HS buffer	1.30 X
MgCl2	2.60 mM
dNTP KAPA	0.26 mM
oIMR1084	0.50 uM
oIMR1085	0.50 uM
oIMR7338	0.50 uM
oIMR7339	0.50 uM
Glycerol	6.50 %
Dye	1.00 X
Kapa 2G HS taq polym	0.03 U/ul
DNA

Cycling

Step	Temp °C	Time	Note
1	94.0	--
2	94.0	--
3	65.0	--	-0.5 C per cycle decrease
4	68.0	--
5		--	repeat steps 2-4 for 10 cycles (Touchdown)
6	94.0	--
7	60.0	--
8	72.0	--
9		--	repeat steps 6-8 for 28 cycles
10	72.0	--
11	10.0	--	hold

JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

JAX uses a 'touchdown' cycling protocol and therefore has not calculated the optimal annealing temperature for each set of primers.

Strains Using This Protocol

Stock Number	Strain Name
005989	129;FVB-Tg(PTH-cre)4167Slib/J
039968	129S-Slc6a4^tm2.1Rbl/JvvwJ
036336	129S.Cg-Ghrhr^tm1.1Deh Trhr^tm1.1Deh/Deh
007179	129S.Cg-Ndor1^Tg(UBC-cre/ERT2)1Ejb/J
007915	129S.FVB-Tg(Amh-cre)8815Reb/J
004302	129S1/Sv-Hprt1^tm1(CAG-cre)Mnn/J
008523	129S6.Cg-Tg(NPHS2-cre)295Lbh/BroJ
039888	B6(129P2)-Ddx3x^em1Tdk/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
022356	B6(129X1)-Tg(Cd4-cre/ERT2)11Gnri/J
039195	B6(Cg)-Il17a^tm1.1(EYFP/icre)Ehs/J
038536	B6(Cg)-Tg(Cd8a-icre/ERT2,-GFP)#Daav/J
026512	B6(Cg)-Tg(TPO-cre/ERT2)1139Tyj/J
038538	B6(Cg)-Tg(Zbtb7b/CD4-icre/ERT2,-CD2)13Daav/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
039931	B6.129(Cg)-Vsx2^tm3.1(cre)Kash/TmjJ
008463	B6.129-Gt(ROSA)26Sor^tm1(cre/ERT2)Tyj/J
025567	B6.129-Olig2^tm1.1(cre)Wdr/J
005697	B6.129-Otx1^tm4(cre)Asim/J
004146	B6.129-Tg(Pcp2-cre)2Mpin/J
008462	B6.129P2-Trp53^tm1Brn/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
036337	B6.Cg-Ghrhr^tm1.1Deh Trhr^tm1.1Deh Tg(CAG-cre/Esr1*)5Amc/Deh
032923	B6.Cg-Kit^tm1(cre)Htng/J
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
038835	B6.Cg-Sdk1^em1(cre/GFP)Jrs/KrishJ
022865	B6.Cg-Trib2^tm1.1(cre/ERT2)Hze/J
039757	B6.Cg-Ucn^tm1.1(cre/EGFP)Rpa/J
017346	B6.Cg-Tg(A930038C07Rik-cre)1Aibs/J
006149	B6.Cg-Tg(ACTA1-cre)79Jme/J
030215	B6.Cg-Tg(Amigo2-cre)1Sieg/J
005359	B6.Cg-Tg(Camk2a-cre)T29-1Stl/J
022071	B6.Cg-Tg(Cd4-cre)1Cwi/BfluJ
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
005069	B6.Cg-Tg(Fabp4-cre)1Rev/J
012712	B6.Cg-Tg(Fev-cre)1Esd/J
035668	B6.Cg-Tg(Gata1-cre)1Sho/MdfJ
009642	B6.Cg-Tg(GH1-cre)1Sac/J
003573	B6.Cg-Tg(Ins2-cre)25Mgn/J
007742	B6.Cg-Tg(Myh11-cre,-EGFP)2Mik/J
010536	B6.Cg-Tg(Pcp2-cre)3555Jdhu/J
041184	B6.Cg-Tg(Pdgfrb-cre)35Vli/Mmjax
005584	B6.Cg-Tg(Prrx1-cre)1Cjt/J
032873	B6.Cg-Tg(Pthlh-cre/ERT2)909Nono/J
040025	B6.Cg-Tg(Pyy-cre)1Able/DbhzJ
003967	B6.Cg-Tg(Rbp3-cre)528Jxm/J
021614	B6.Cg-Tg(S100A8-cre,-EGFP)1Ilw/J
017491	B6.Cg-Tg(Tagln-cre)1Her/J
004128	B6.Cg-Tg(Tek-cre)12Flv/J
008863	B6.Cg-Tg(Tek-cre)1Ywa/J
006234	B6.Cg-Tg(tetO-cre)1Jaw/J
035670	B6.Cg-Tg(VAV1-cre)1Graf/MdfJ
020282	B6.Cg-Tg(Vil1-cre/ERT2)23Syr/J
006451	B6.FVB(129X1)-Tg(Sim1-cre)1Lowl/J
039769	B6.FVB(C)-Tg(Pdpn-cre)HRkl/J
006333	B6.FVB(Cg)-Tg(Neurog3-cre)C1Able/J
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
003394	B6.FVB-Tg(Zp3-cre)3Mrt/J
033052	B6;129-Procr^tm1(cre/ERT2)Yaz/J
009600	B6;129-Six2^tm3(EGFP/cre/ERT2)Amc/J
024860	B6;129-Tg(Drd1-cre)120Mxu/Mmjax
017593	B6;129S-Sox2^tm1(cre/ERT2)Hoch/J
036300	B6;129S4-Cux1^tm1(cre/ERT2)Zjh/J
012362	B6;129S6-Tg(Camk2a-cre/ERT2)1Aibs/J
017495	B6;129S7-Crim1^tm1(GFP/cre/ERT2)Pzg/J
038989	B6;C3-Oxtr^em4(icre/ERT2)Yinn/2J
009616	B6;C3-Tg(A930038C07Rik-cre)4Aibs/J
008844	B6;C3-Tg(Ctgf-cre)2Aibs/J
008839	B6;C3-Tg(Cyp39a1-cre)1Aibs/J
009117	B6;C3-Tg(Cyp39a1-cre)7Aibs/J
009111	B6;C3-Tg(Scnn1a-cre)1Aibs/J
009103	B6;C3-Tg(Wfs1-cre/ERT2)3Aibs/J
025809	B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J
003466	B6;D2-Tg(Sycp1-cre)4Min/J
015855	B6;DBA-Tg(Upk3a-GFP/cre/ERT2)26Amc/J
023748	B6;FVB-Tg(Aldh1l1-cre)JD1884Htz/J
003734	B6;FVB-Tg(GZMB-cre)1Jcb/J
004426	B6;SJL-Tg(Cga-cre)3Sac/J
003554	B6;SJL-Tg(Col2a1-cre)1Bhr/J
017738	B6;SJL-Tg(Foxl1-cre)1Khk/J
005249	B6;SJL-Tg(Krt1-15-cre/PGR*)22Cot/J
003552	B6129-Tg(Wap-cre)11738Mam/J
007252	B6Ei.129S4-Tg(Prm-cre)58Og/EiJ
040426	B6J(B6N)-Tg(Nr4a1-EGFP/cre)820Khog/PalcaJ
018966	B6N.Cg-Tg(Camk2a-cre)T29-1Stl/J
018965	B6N.Cg-Tg(Fabp4-cre)1Rev/J
018960	B6N.Cg-Tg(Ins2-cre)25Mgn/J
018964	B6N.Cg-Tg(KRT14-cre)1Amc/J
003465	BALB/c-Tg(CMV-cre)1Cgn/J
004126	C.Cg-Cd19^tm1(cre)Cgn Igh^b/J
005673	C.Cg-Tg(Mx1-cre)1Cgn/J
006244	C.Cg-Tg(tetO-cre)1Jaw/J
039447	C.FVB-Tg(Pdpn-cre)HRkl/J
040186	C57BL/6-Adora2a^em1(cre)Rcos/J
040185	C57BL/6-Anxa1^em1(cre)Rcos/J
040715	C57BL/6-Avpr1a^tm1.1Gdol/J
040188	C57BL/6-Drd1^em1(cre)Rcos/J
035489	C57BL/6-Elmo1^tm1Mki/J
038053	C57BL/6-Oprm1^tm2.1(EGFP/cre)Ics/KffJ
016097	C57BL/6-Tg(Car1-cre)5Flt/J
026828	C57BL/6-Tg(Cpa3-cre)4Glli/J
040040	C57BL/6-Tg(Fap-cre)1Rkl/J
006474	C57BL/6-Tg(Grik4-cre)G32-4Stl/J
008870	C57BL/6-Tg(Hspa2-cre)1Eddy/J
016261	C57BL/6-Tg(Nes-cre/ERT2)KEisc/J
013148	C57BL/6-Tg(Pdgfra-cre)1Clc/J
032131	C57BL/6-Tg(Scx-GFP/cre)1Stzr/J
040483	C57BL/6J-Agrp^em1(cre)Kmso/J
040493	C57BL/6J-Cst7^em3(folA/cre)Aduci/J
040192	C57BL/6J-Ecel1^em1(cre)Gfng/J
041040	C57BL/6J-Enpp6^em1(cre/ERT2)Lusu/J
036429	C57BL/6J-Gne^tm1Dar/J
040885	C57BL/6J-Lmx1b^em1(cre)Jgeer/J
039261	C57BL/6J-Ntrk2^em1(cre)Bxu/J
040191	C57BL/6J-Spp1^em1(cre)Gfng/J
032080	C57BL/6J-Tg(Cd8a*-cre)B8Asin/J
003651	C57BL/6J-Tg(Zp3-cre)93Knw/J
027789	C57BL/6N-Fgd5^tm3(cre/ERT2)Djr/J
040961	C57BL/6N-Ighg1^em2(cre,-dre/ERT2)Rsky/J
023453	C57BL/6N-Lhx8^tm1(cre)Grsr/2GrsrJ
012686	C57BL/6N-Tg(Ppp1r2-cre)4127Nkza/J
012929	FVB(Cg)-Tg(Dhh-cre)1Mejr/J
005025	FVB.Cg-Grm7^Tg(SMN2)89Ahmb Smn1^tm1Msd Tg(SMN2*delta7)4299Ahmb/J
006139	FVB.Cg-Tg(ACTA1-cre)79Jme/J
017595	FVB.Cg-Tg(CAG-cre/Esr1*)5Amc/J
006297	FVB.Cg-Tg(Eno2-cre)39Jme/J
018394	FVB.Cg-Tg(KRT5-cre/ERT2)2Ipc/JeldJ
008244	FVB.Cg-Tg(tetO-cre)1Jaw/J
003314	FVB/N-Tg(EIIa-cre)C5379Lmgd/J
006143	FVB/N-Tg(Thy1-cre)1Vln/J
003377	FVB/N-Tg(Zp3-cre)3Mrt/J
013233	NOD.B6-Tg(Itgax-cre,-EGFP)4097Ach/J
034859	NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg(CAG-cre/Esr1*)5Amc/J
004986	NOD/ShiLt-Tg(Ins2-cre)3Lt/LtJ
003855	NOD/ShiLt-Tg(Ins2-cre)5Lt/LtJ
035185	STOCK Adgrl1^tm2c(EUCOMM)Hmgu/SudJ
036942	STOCK Ager^tm2.1(cre/ERT2)Blh/2J
039535	STOCK Aldh1a3^tm1(cre/ERT2)Dac/J
037028	STOCK Calcr^tm1.1(cre)Mgmj/J
039985	STOCK Cysltr2^em1(cre)Ddg/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
018418	STOCK Lrig1^tm1.1(cre/ERT2)Rjc/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
039729	STOCK Pnmt^tm1.1(cre)Sne/J
040319	STOCK Slc34a1^em1(EGFP/cre)Jsprk/J
020080	STOCK Slc6a3^tm1(cre)Xz/J
036906	STOCK Tmem178b^tm1c(KOMP)Wtsi Tmem178^tm1.1Sud/J
039435	STOCK Tnc^tm1.1(cre/ERT2)Gcr/J
010911	STOCK Wt1^tm1(EGFP/cre)Wtp/J
007684	STOCK Tg(Atoh1-cre/Esr1*)14Fsh/J
032910	STOCK Tg(BEST1-rtTA,tetO-cre)1Yzl/J
004453	STOCK Tg(CAG-cre/Esr1*)5Amc/J
035566	STOCK Tg(CAG-mRuby2*,-mClover*)1Srgt/J
009615	STOCK Tg(Cartpt-cre)1Aibs/J
017336	STOCK Tg(Cd4-cre)1Cwi/BfluJ
025736	STOCK Tg(Cela1-cre/ERT)1Lgdn/J
005105	STOCK Tg(Chx10-EGFP/cre,-ALPP)2Clc/J
008852	STOCK Tg(En2-cre)22Alj/J
005938	STOCK Tg(Eno2-cre)39Jme/J
022763	STOCK Tg(Eno2-cre/ERT2)1Pohlk/J
012841	STOCK Tg(Ggt1-cre)M3Egn/J
027681	STOCK Tg(Hnf1b-cre/ERT2)1Jfer/J
004782	STOCK Tg(KRT14-cre)1Amc/J
005107	STOCK Tg(KRT14-cre/ERT)20Efu/J
017948	STOCK Tg(KRT18-cre/ERT2)23Blpn/J
023724	STOCK Tg(mI56i-cre,-EGFP)1Kc/J
003553	STOCK Tg(MMTV-cre)4Mam/J
008847	STOCK Tg(Msx2-cre/ERT)27Alj/J
009074	STOCK Tg(Myh6-cre)1Jmk/J
028364	STOCK Tg(Neurod1-cre)1Able/J
005667	STOCK Tg(Neurog3-cre)C1Able/J
006207	STOCK Tg(Pcp2-cre)1Amc/J
005965	STOCK Tg(Pomc1-cre)16Lowl/J
018829	STOCK Tg(Sox9-cre/ERT2)1Msan/J
032646	STOCK Tg(Spo11-cre)1Rsw/PecoJ
004746	STOCK Tg(Tagln-cre)1Her/J
006224	STOCK Tg(tetO-cre)1Jaw/J
008851	STOCK Tg(Wnt1-cre/ERT)1Alj/J
200 strains use this protocol