Protocol 20626: Probe Assay - Generic Cre Probe
Version 3.0

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 Tm = 84°C +/- 1.0°C
Internal positive control Tm = 79.4°C+/- 1.0°C

Transgene = 102 bp
Internal Positive Control = 74 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
oIMR1084 GCG GTC TGG CAG TAA AAA CTA TC Transgene Forward A Cre
oIMR1085 GTG AAA CAG CAT TGC TGT CAC TT Transgene Reverse A Cre
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
005989 129;FVB-Tg(PTH-cre)4167Slib/J
017789 129S-Slc11a2tm2Nca/J
007179 129S.Cg-Ndor1Tg(UBC-cre/ERT2)1Ejb/J
007915 129S.FVB-Tg(Amh-cre)8815Reb/J
004302 129S1/Sv-Hprt1tm1(CAG-cre)Mnn/J
008523 129S6.Cg-Tg(NPHS2-cre)295Lbh/BroJ
030182 B6(129S4)-Foxc2tm1.1(cre/ERT2)Rssr/J
033169 B6(129S4)-Kiss1tm1.1(cre/EGFP)Rpa/J
032051 B6(129S4)-Kittm1.1(cre)Jmol/J
037511 B6(129S4)-Pou2f3tm1.1(cre/ERT2)Imt/J
009575 B6(129S4)-Et(cre/ERT2)119Rdav/J
009580 B6(129S4)-Et(cre/ERT2)1382Rdav/J
009581 B6(129S4)-Et(cre/ERT2)1642Rdav/J
009582 B6(129S4)-Et(cre/ERT2)1645Rdav/J
009583 B6(129S4)-Et(cre/ERT2)1957Rdav/J
009584 B6(129S4)-Et(cre/ERT2)2007Rdav/J
009585 B6(129S4)-Et(cre/ERT2)2047Rdav/J
009574 B6(129S4)-Et(cre/ERT2)21Rdav/J
009577 B6(129S4)-Et(cre/ERT2)296Rdav/J
009578 B6(129S4)-Et(cre/ERT2)398Rdav/J
009573 B6(129S4)-Et(cre/ERT2)4Rdav/J
009579 B6(129S4)-Et(cre/ERT2)837Rdav/J
029290 B6(Cg)-Sgtatm1.1Drmcr/J
021072 B6(Cg)-Slc6a8tm1.2Clar/J
033895 B6(Cg)-Tg(Snap25-FLPe)47DSJDJdd Tg(Egr1-cre/ERT2)47DSJDJdd/J
026512 B6(Cg)-Tg(TPO-cre/ERT2)1139Tyj/J
021012 B6(D2)-Tg(CAG-Brainbow1.0)3Eggn/J
037052 B6(FVB)-Tg(Sprr2f-cre)2Dcas/J
021881 B6.129(Cg)-Arctm1.1(cre/ERT2)Luo/J
018057 B6.129(Cg)-Cul7tm2.1Jdec/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
029228 B6.129-Bhlha15tm3(cre/ERT2)Skz/J
008463 B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J
018445 B6.129-Mre11atm1.1Rchd/J
025567 B6.129-Olig2tm1.1(cre)Wdr/J
005697 B6.129-Otx1tm4(cre)Asim/J
004146 B6.129-Tg(Pcp2-cre)2Mpin/J
021160 B6.129P2(Cg)-Cx3cr1tm2.1(cre/ERT2)Litt/WganJ
018054 B6.129P2(Cg)-Pitx2tm1.1Dmm/J
023726 B6.129P2-C1qatm1.1Tenn/J
016934 B6.129P2-Lgr6tm2.1(cre/ERT2)Cle/J
018997 B6.129S(FVB)-Lhx3tm1.1Sjr/J
032621 B6.129S-Ly6atm1.1(cre)Jmol/J
016132 B6.129S4-Perptm2Att/J
006447 B6.129S6(CBA)-Cebpatm1Dgt/J
012900 B6.129S6-Stat5btm1Mam Stat5atm2Mam/Mmjax
025929 B6.129S7(Cg)-Pik3catm1Jdo/J
006054 B6.C-Tg(CMV-cre)1Cgn/J
030644 B6.C-Tg(S100a4-cre)1Egn/JhrsJ
008601 B6.Cg-7630403G23RikTg(Th-cre)1Tmd/J
006230 B6.Cg-Cebpatm1Dgt Tg(Mx1-cre)1Cgn/J
024521 B6.Cg-Eif4g1tm1.2Mjff/J
036450 B6.Cg-Foxl1tm2(cre/ERT2)Khk/Mmjax
032923 B6.Cg-Kittm1(cre)Htng/J
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
030546 B6.Cg-Satb2tm1.1(cre/GFP)Rpa/J
017889 B6.Cg-Shank3tm1.1Bux/J
032488 B6.Cg-Six2tm2(cre/ERT2)Amc/J
022865 B6.Cg-Trib2tm1.1(cre/ERT2)Hze/J
028538 B6.Cg-Zbtb46tm3.1(cre)Mnz/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
022357 B6.Cg-Tg(Arc-cre/ERT2)MRhn/CdnyJ
027400 B6.Cg-Tg(Camk2a-cre)3Szi/J
009352 B6.Cg-Tg(CDX2-cre*)189Erf/J
016241 B6.Cg-Tg(Col1a1-cre/ERT2)1Crm/J
016237 B6.Cg-Tg(Col1a2-cre/ERT,-ALPP)7Cpd/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
009642 B6.Cg-Tg(GH1-cre)1Sac/J
003573 B6.Cg-Tg(Ins2-cre)25Mgn/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
004128 B6.Cg-Tg(Tek-cre)12Flv/J
008863 B6.Cg-Tg(Tek-cre)1Ywa/J
012328 B6.Cg-Tg(Tyr-cre/ERT2)13Bos/J
035670 B6.Cg-Tg(VAV1-cre)1Graf/MdfJ
020282 B6.Cg-Tg(Vil1-cre/ERT2)23Syr/J
028353 B6.Cg-Tg(Vil1-EGFP)106Gum/J
024688 B6.FVB(129S)-Tg(Pax6-GFP/cre)1Rilm/J
006451 B6.FVB(129X1)-Tg(Sim1-cre)1Lowl/J
028736 B6.FVB(Cg)-Tg(Mylpf-cre)3Kraj/Mmjax
028020 B6.FVB-Tg(Adipoq-cre)1Evdr/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
011069 B6.FVB-Tg(Gh1-cre)bKnmn/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
017601 B6;129-Eif2s1tm1Rjk/J
016210 B6;129-Lrrk2tm3.1Shn/J
018319 B6;129-Plxnd1tm1.1Tmj/J
009600 B6;129-Six2tm3(EGFP/cre/ERT2)Amc/J
018773 B6;129-Sult2b1tm1Rus/J
024860 B6;129-Tg(Drd1-cre)120Mxu/Mmjax
002914 B6;129S-Hexbtm1Rlp/J
022864 B6;129S-Rasgrf2tm1(cre/folA)Hze/J
017593 B6;129S-Sox2tm1(cre/ERT2)Hoch/J
009388 B6;129S1-Osr2tm2(cre)Jian/J
036300 B6;129S4-Cux1tm1(cre/ERT2)Zjh/J
018019 B6;129S4-Srsf2tm1Xdfu/J
009576 B6;129S4-Et(cre/ERT2)278Rdav/J
017495 B6;129S7-Crim1tm1(GFP/cre/ERT2)Pzg/J
018937 B6;C-Tg(Erv4-cre/ERT2*)657Slp/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
009112 B6;C3-Tg(Scnn1a-cre)2Aibs/J
009103 B6;C3-Tg(Wfs1-cre/ERT2)3Aibs/J
031779 B6;CBA-Tg(Cyp19a1-cre)1Dlc/J
025809 B6;CBA-Tg(Fgfr3-icre/ERT2)4-2Wdr/J
024926 B6;D2-Tg(Fshr-cre)1Ldu/J
003466 B6;D2-Tg(Sycp1-cre)4Min/J
015855 B6;DBA-Tg(Upk3a-GFP/cre/ERT2)26Amc/J
018402 B6;FVB-Nrn1tm1.2Ndiv/J
023748 B6;FVB-Tg(Aldh1l1-cre)JD1884Htz/J
003734 B6;FVB-Tg(GZMB-cre)1Jcb/J
026526 B6;SJL-Akap6tm1.1Mskf/Mmjax
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
017704 B6N.129-Pik3catm1Jjz/J
017911 B6N.129S6(Cg)-Esr1tm1.1(cre)And/J
016881 B6N.Cg-Adgra2tm1.1Bstc/J
019102 B6N.Cg-Tg(CAG-cre/Esr1*)5Amc/CjDswJ
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
019103 B6N.Cg-Tg(Nes-cre)1Kln/CjDswJ
019099 B6N.FVB-Tmem163Tg(ACTB-cre)2Mrt/CjDswJ
030894 B6NCrl.129P2(B6)-Apoetm1Unc Ccr5tm1Blck/BlckJ
003465 BALB/c-Tg(CMV-cre)1Cgn/J
013756 C.129S-Batf3tm1Kmm/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
009155 C57BL/6-Cldn6tm1(cre)Dkwu/J
035489 C57BL/6-Elmo1tm1Mki/J
017557 C57BL/6-Tg(BEST1-cre)1Jdun/J
016097 C57BL/6-Tg(Car1-cre)5Flt/J
026828 C57BL/6-Tg(Cpa3-cre)4Glli/J
008870 C57BL/6-Tg(Hspa2-cre)1Eddy/J
024034 C57BL/6-Tg(Pmch-cre)1Rck/J
036429 C57BL/6J-Gnetm1Dar/J
030379 C57BL/6J-Tg(Abcb1a-cre)NIDA045Htz/J
032080 C57BL/6J-Tg(Cd8a*-cre)B8Asin/J
018899 C57BL/6J-Tg(Krt6,-cre,-Cerulean)5Grsr/Grsr
021582 C57BL/6J-Tg(Mchr1-cre)1Emf/J
022887 C57BL/6J-Tg(Six6-cre)7Grsr/GrsrJ
027789 C57BL/6N-Fgd5tm3(cre/ERT2)Djr/J
023453 C57BL/6N-Lhx8tm1(cre)Grsr/2GrsrJ
038053 C57BL/6N-Oprm1tm2.1(EGFP/cre)Ics/KffJ
030484 C57BL/6N-Tg(Oxr1-cre)C14Stl/J
012686 C57BL/6N-Tg(Ppp1r2-cre)4127Nkza/J
028325 C57BL/6NJ-Cldn13em2J/J
026266 D2.B6-Tg(Zp3-cre)93Knw/SjJ
026852 D2.Cg-Tg(Gfap-cre)73.12Mvs/SjJ
026860 D2.FVB-Tg(Tek-cre)2352Rwng/SjJ
012929 FVB(Cg)-Tg(Dhh-cre)1Mejr/J
004600 FVB-Tg(GFAP-cre)25Mes/J
005025 FVB.Cg-Grm7Tg(SMN2)89Ahmb Smn1tm1Msd 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
008244 FVB.Cg-Tg(tetO-cre)1Jaw/J
003314 FVB/N-Tg(EIIa-cre)C5379Lmgd/J
025066 FVB/N-Tg(Mylpf-cre)3Kraj/Mmjax
003377 FVB/N-Tg(Zp3-cre)3Mrt/J
017678 FVB;129-Pink1tm1Aub Tg(Prnp-SNCA*A53T)AAub/J
023325 FVB;B6-Tg(Pbsn-cre)20Fwan/J
034859 NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg(CAG-cre/Esr1*)5Amc/J
034795 NOD.Cg-Tg(CAG-cre/Esr1*)5Amc/J
013251 NOD.FVB-Tg(EIIa-cre)C5379Lmgd/J
004986 NOD/ShiLt-Tg(Ins2-cre)3Lt/LtJ
003855 NOD/ShiLt-Tg(Ins2-cre)5Lt/LtJ
035185 STOCK Adgrl1tm2c(EUCOMM)Hmgu/SudJ
036942 STOCK Agertm2.1(cre/ERT2)Blh/2J
012899 STOCK Agrptm1(cre)Lowl/J
025665 STOCK Asxl1tm1.1Iaai/J
037028 STOCK Calcrtm1.1(cre)Mgmj/J
007916 STOCK En1tm2(cre)Wrst/J
026199 STOCK F8tm1.1Rmnt/J
027012 STOCK Foxj1tm1.1(cre/ERT2/GFP)Htg/J
007913 STOCK Gli1tm3(cre/ERT2)Alj/J
008783 STOCK Grm7Tg(SMN2)89Ahmb Smn1tm3(SMN2/Smn1)Mrph Tg(CAG-cre/Esr1*)5Amc Tg(SMN2*delta7)4299Ahmb/J
022793 STOCK Gt(ROSA)26Sortm1(LRRK2*R1441C)Djmo/J
024283 STOCK Hcn4tm2.1(cre/ERT2)Sev/J
017298 STOCK Hmgn5tm1.1Mbus/J
035978 STOCK Hoxb8tm2.1(cre)Mrc/J
032429 STOCK Krastm4Tyj Trp53tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
018418 STOCK Lrig1tm1.1(cre/ERT2)Rjc/J
016945 STOCK Mir138-1tm1Mtm/Mmjax
004192 STOCK Mttptm2Sgy Ldlrtm1Her Apobtm2Sgy Tg(Mx1-cre)1Cgn/J
025669 STOCK Myf6tm1.1(cre/ERT2)Lepr/J
030291 STOCK Ntrk3tm3.1(cre/ERT2)Ddg/J
035303 STOCK Obscntm1Chen/J
035925 STOCK Opn4tm1(cre)Saha/J
035045 STOCK Oprk1tm1.1(cre)Sros/J
030543 STOCK Oxtrtm1(cre/GFP)Rpa/J
022972 STOCK Pros1tm1Grl/J
023329 STOCK Ptf1atm1(cre)Hnak/RschJ
019378 STOCK Ptf1atm2(cre/ESR1)Cvw/J
013086 STOCK Rxratm1Krc/J
020080 STOCK Slc6a3tm1(cre)Xz/J
023405 STOCK Tfap2atm1Hsv/J
036906 STOCK Tmem178btm1c(KOMP)Wtsi Tmem178tm1.1Sud/J
010911 STOCK Wt1tm1(EGFP/cre)Wtp/J
024849 STOCK Et(cre)TFC.09Lois/J
025750 STOCK Tg(ACTA1-cre/Esr1*)2Kesr/J
027826 STOCK Tg(Actl6b-Cre)4092Jiwu/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
031049 STOCK Tg(Colgalt2-cre)NF111Htz/J
008852 STOCK Tg(En2-cre)22Alj/J
005938 STOCK Tg(Eno2-cre)39Jme/J
022763 STOCK Tg(Eno2-cre/ERT2)1Pohlk/J
026625 STOCK Tg(Gcm1-cre)1Chrn/J
029485 STOCK Tg(Gsx1-cre)6Wql/J
027681 STOCK Tg(Hnf1b-cre/ERT2)1Jfer/J
019380 STOCK Tg(IVL-cre/ERT2)1Blpn/J
029417 STOCK Tg(Jam2-cre/ERT2)2Jrs/J
008582 STOCK Tg(Kcnc2-Cre)K128Stl/LetJ
005107 STOCK Tg(KRT14-cre/ERT)20Efu/J
017948 STOCK Tg(KRT18-cre/ERT2)23Blpn/J
009643 STOCK Tg(Lhb-cre)1Sac/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
009102 STOCK Tg(Nefh-cre)12Kul/J
028364 STOCK Tg(Neurod1-cre)1Able/J
005667 STOCK Tg(Neurog3-cre)C1Able/J
006207 STOCK Tg(Pcp2-cre)1Amc/J
009606 STOCK Tg(Six2-EGFP/cre)1Amc/J
032646 STOCK Tg(Spo11-cre)1Rsw/PecoJ
006224 STOCK Tg(tetO-cre)1Jaw/J
008851 STOCK Tg(Wnt1-cre/ERT)1Alj/J
018281 STOCK Tg(Wnt7a-EGFP/cre)#Bhr/Mmjax
259 strains use this protocol