Stock No: 008870
Protocol 22392: Standard PCR Assay - Generic Cre
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
007179 129S.Cg-Ndor1Tg(UBC-cre/ERT2)1Ejb/J
007915 129S.FVB-Tg(Amh-cre)8815Reb/J
008523 129S6.Cg-Tg(NPHS2-cre)295Lbh/BroJ
008320 B6.129(Cg)-Leprtm2(cre)Rck/J
005697 B6.129-Otx1tm4(cre)Asim/J
004146 B6.129-Tg(Pcp2-cre)2Mpin/J
006600 B6.129S1-Mnx1tm4(cre)Tmj/J
006054 B6.C-Tg(CMV-cre)1Cgn/J
008601 B6.Cg-7630403G23RikTg(Th-cre)1Tmd/J
006230 B6.Cg-Cebpatm1Dgt Tg(Mx1-cre)1Cgn/J
022865 B6.Cg-Trib2tm1.1(cre/ERT2)Hze/J
017346 B6.Cg-Tg(A930038C07Rik-cre)1Aibs/J
006149 B6.Cg-Tg(ACTA1-cre)79Jme/J
005359 B6.Cg-Tg(Camk2a-cre)T29-1Stl/J
006368 B6.Cg-Tg(Cr2-cre)3Cgn/J
008538 B6.Cg-Tg(Cspg4-cre/Esr1*)BAkik/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
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
005584 B6.Cg-Tg(Prrx1-cre)1Cjt/J
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
008863 B6.Cg-Tg(Tek-cre)1Ywa/J
006234 B6.Cg-Tg(tetO-cre)1Jaw/J
006451 B6.FVB(129X1)-Tg(Sim1-cre)1Lowl/J
006333 B6.FVB(Cg)-Tg(Neurog3-cre)C1Able/J
006137 B6.FVB-Tg(Cdh5-cre)7Mlia/J
003724 B6.FVB-Tg(EIIa-cre)C5379Lmgd/J
010714 B6.FVB-Tg(Pomc-cre)1Lowl/J
003394 B6.FVB-Tg(Zp3-cre)3Mrt/J
017593 B6;129S-Sox2tm1(cre/ERT2)Hoch/J
017495 B6;129S7-Crim1tm1(GFP/cre/ERT2)Pzg/J
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
003466 B6;D2-Tg(Sycp1-cre)4Min/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
007252 B6Ei.129S4-Tg(Prm-cre)58Og/EiJ
018966 B6N.Cg-Tg(Camk2a-cre)T29-1Stl/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-Cd19tm1(cre)Cgn Ighb/J
005673 C.Cg-Tg(Mx1-cre)1Cgn/J
006244 C.Cg-Tg(tetO-cre)1Jaw/J
016097 C57BL/6-Tg(Car1-cre)5Flt/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
003651 C57BL/6-Tg(Zp3-cre)93Knw/J
034448 C57BL/6J-Tlr9em1Ldm/J
032080 C57BL/6J-Tg(Cd8a*-cre)B8Asin/J
027789 C57BL/6N-Fgd5tm3(cre/ERT2)Djr/J
012686 C57BL/6N-Tg(Ppp1r2-cre)4127Nkza/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
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
004986 NOD/ShiLt-Tg(Ins2-cre)3Lt/LtJ
003855 NOD/ShiLt-Tg(Ins2-cre)5Lt/LtJ
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
008783 STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J
032429 STOCK Krastm4Tyj Trp53tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
035045 STOCK Oprk1tm1.1(cre)Sros/J
007684 STOCK Tg(Atoh1-cre/Esr1*)14Fsh/J
004453 STOCK Tg(CAG-cre/Esr1*)5Amc/J
009615 STOCK Tg(Cartpt-cre)1Aibs/J
017336 STOCK Tg(Cd4-cre)1Cwi/BfluJ
005105 STOCK Tg(Chx10-EGFP/cre,-ALPP)2Clc/J
008852 STOCK Tg(En2-cre)22Alj/J
005938 STOCK Tg(Eno2-cre)39Jme/J
012841 STOCK Tg(Ggt1-cre)M3Egn/J
004782 STOCK Tg(KRT14-cre)1Amc/J
017948 STOCK Tg(KRT18-cre/ERT2)23Blpn/J
017947 STOCK Tg(Krt8-cre/ERT2)17Blpn/J
003553 STOCK Tg(MMTV-cre)4Mam/J
008847 STOCK Tg(Msx2-cre/ERT)27Alj/J
009074 STOCK Tg(Myh6-cre)1Jmk/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
104 strains use this protocol