Protocol 27167: Standard PCR Assay - Generic Cre/ERT2 alternate1
Version 1.2

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 = ~200 bp

Internal positive control = 324 bp

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
19974 GGC CAG GCT GTT CTT CTT AG Transgene Reverse A ERT2
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
oIMR8346 ATA CCG GAG ATC ATG CAA GC Transgene Forward A Cre

Reaction A

Component Final Concentration
ddH2O
Kapa 2G HS buffer 1.30 X
MgCl2 2.60 mM
dNTP KAPA 0.26 mM
19974 0.50 uM
oIMR7338 0.50 uM
oIMR7339 0.50 uM
oIMR8346 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
039977 129S.Cg-Tg(Arc-cre/ERT2)MRhn/CdnyJ
037511 B6(129S4)-Pou2f3tm1.1(cre/ERT2)Imt/J
032132 B6(129S4)-Scxtm2.1(cre/ERT2)Stzr/J
041008 B6(129S6)-Scube2em1(cre/ERT2)Tsuki/J
038514 B6(129S6)-Tg(Slc6a5-cre/ERT2)#Uze/J
040559 B6(Cg)-Cxcr4tm1.1(cre/ERT2)Stum/J
038595 B6(Cg)-Dcxem1(cre/ERT2)Jjan/J
026802 B6(Cg)-Ins1tm2.1(cre/ERT2)Thor/J
032758 B6(Cg)-Tg(Acta2-cre/ERT2)1Ikal/J
039060 B6.129S2-Albtm1(cre/ERT2)Mtz/J
038495 B6.129S6(Cg)-Gli1tm3(cre/ERT2)Alj/RtigJ
038129 B6.Cg-Ascl3tm2.1(icre/ESR1*)Ovi/J
037804 B6.Cg-Clec9atm4.1(cre/ERT2)Crs/J
036450 B6.Cg-Foxl1tm2(cre/ERT2)Khk/Mmjax
035169 B6.Cg-Krastm4Tyj Apctm1Tno Tg(CDX2-cre/ERT2)752Erf/MaraJ
038956 B6.Cg-Npr3tm2(cre/ERT2)Hze/J
039726 B6.Cg-Sall4tm1.1(EGFP/cre/ERT2)Ykawa/J
038834 B6.Cg-Sdk1tm1.1(cre/ERT2)Jrs/KrishJ
038840 B6.Cg-Sdk2tm1.1(cre/ERT2)Jrs/KrishJ
039530 B6.Cg-Sema6atm1.1(cre/ERT2)Alk/J
041033 B6.Cg-Tnnt1tm1.1(cre/ERT2)Hze/J
029567 B6.Cg-Tg(Col1a2-cre/ERT,-ALPP)7Cpd/2J
016237 B6.Cg-Tg(Col1a2-cre/ERT,-ALPP)7Cpd/J
039049 B6.Cg-Tg(Col2a1-cre)3Amc/RgrayJ
032757 B6.Cg-Tg(Dmp1-cre/ERT2)0022Pdp/IkalJ
012849 B6.Cg-Tg(GFAP-cre/ERT2)505Fmv/J
024709 B6.Cg-Tg(Ins1-cre/ERT)1Lphi/J
038390 B6.Cg-Tg(KRT14-cre/ERT2)1Ipc/MtzJ
029684 B6.Cg-Tg(Pdgfrb-cre/ERT2)6096Rha/J
005975 B6.Cg-Tg(Plp1-cre/ERT)3Pop/J
029211 B6.Cg-Tg(Prrx1-cre/ERT2,-EGFP)1Smkm/J
013590 B6.Cg-Tg(Tyr-cre/ERT2)13Bos Braftm1Mmcm Ptentm1Hwu/BosJ
020282 B6.Cg-Tg(Vil1-cre/ERT2)23Syr/J
029231 B6;129-Bhlha15tm4Skz/J
039579 B6;129-Eya1tm3(cre/ERT2)Px/J
036292 B6;129-Gt(ROSA)26Sortm1(BEST1-cre/ERT2)Vold/J
036300 B6;129S4-Cux1tm1(cre/ERT2)Zjh/J
036301 B6;129S4-Eomestm1.1(cre/ERT2)Zjh/J
036296 B6;129S4-Fezf2tm1.1(cre/ERT2)Zjh/J
030346 B6;129S4-Gcgem1(cre/ERT2)Khk/Mmjax
036298 B6;129S4-Tle4tm1.1(cre/ERT2)Zjh/J
040784 B6;129S6-Pgrem1(icre/ERT2)Fjd/J
012362 B6;129S6-Tg(Camk2a-cre/ERT2)1Aibs/J
014160 B6;DBA-Tg(S100b-EGFP/cre/ERT2)22Amc/J
035554 B6;FVB-Tg(Lfng-cre/ERT2)1Mmsa/J
036751 B6;SJL-Gfralem2(cre/ERT2)Rsy/J
036770 B6;SJL-Tg(Gm5127-cre/ERT2)1Mhsi/J
039615 B6129S1-Emx2em1(cre/ERT2)Mdea/J
037087 B6J.Cg-Tg(Mnx1-cre/ERT2)8Rbro/Mmjax
018280 B6N.Cg-Tg(Pdgfra-cre/ERT)467Dbe/J
033019 B6N.Cg-Tg(Vil1-cre)20Syr/J
031008 B6N.FVB-Tg(Aldh1l1-cre/ERT2)1Khakh/J
038602 C57BL/6-Car4em1(cre/ERT2)Jich/J
040586 C57BL/6-Cntn2em1(cre/ERT2)Forni/J
038831 C57BL/6-Efnb2em1(cre/ERT2)Kloh/J
041040 C57BL/6-Enpp6em1(cre/ERT2)Lusu/J
040119 C57BL/6-Lyve1em1(cre/ERT2)Jkpns/J
040118 C57BL/6-Mrc1em1(cre/ERT2)Jkpns/J
035973 C57BL/6-Rpe65tm1.1(cre/ERT2)Kser/J
038640 C57BL/6-Rtkn2em1(cre/ERT2)Jich/J
041115 C57BL/6-Wif1em1(cre/ERT2)Tchn/J
036499 C57BL/6-Tg(Aire-cre/Esr1)1Mmat/J
034244 C57BL/6-Tg(Atp4b-cre/ERT2)1Jrgo/J
037658 C57BL/6-Tg(Myh11-cre/ERT2)F31Gko/J
036217 C57BL/6-Tg(Nanog-cre/Esr1*)4470Mms/Mmjax
016261 C57BL/6-Tg(Nes-cre/ERT2)KEisc/J
037467 C57BL/6-Tg(Tal1-cre/ERT)1Jrg/J
037466 C57BL/6-Tg(Tal1-cre/ERT)42-056Jrg/J
037483 C57BL/6J-Atf3em1(cre/ERT2)Wlf/J
039262 C57BL/6J-Cthrc1em1(cre/ERT2)Rph/J
039217 C57BL/6J-Dpp4em1(cre/ERT2)Jkpns/J
040457 C57BL/6J-Gprc5aem1(icre/ERT2)Blr/Mmjax
038824 C57BL/6J-Myh11em2(cre/ERT2)Jifz/J
039218 C57BL/6J-Slc47a1em1(cre/ERT2)Jkpns/J
037591 C57BL/6J-Tg(Dct-cre/ERT2)40ALru/J
036482 C57BL/6J-Tg(Gata1-cre/ERT2)L259Jeng/J
037269 C57BL/6J-Tg(Krt8-cre/ERT2,-GFP)1Xin/J
036976 FVB.129S6(Cg)-Krt5tm1.1(cre/ERT2)Blh/MqrJ
035703 FVB.B6-Igfbp2em1(cre/ERT2)Ntang/J
024760 FVB/N-Tg(Ddx4-cre/ERT2)1Dcas/J
036735 STOCK Adgra3tm1.1(HBEGF,-cre/ERT2)Pac/Mmjax
039560 STOCK Adra2atm1(cre/ERT2)Ddg/J
036942 STOCK Agertm2.1(cre/ERT2)Blh/2J
036343 STOCK Apoc4em1(cre/ERT2)Hzhu/J
036246 STOCK Cyp1a2em1(cre/ERT2)Hzhu/J
038582 STOCK Fbxo2tm1(cre/ERT2)Akg/J
038587 STOCK Foxi3em2(cre/ERT2)Akg/J
036247 STOCK Gls2em1(cre/ERT2)Hzhu/J
036248 STOCK Glulem1(cre/ERT2)Hzhu/J
039021 STOCK Gpr83tm1.1(cre/ERT2)Ddg/J
024283 STOCK Hcn4tm2.1(cre/ERT2)Sev/J
041110 STOCK Kmt2ctm1.2Jwle Gt(ROSA)26Sortm1(cre/ERT2)Tyj Kmt2dem2Kaig/KaigMmjax
032429 STOCK Krastm4Tyj Trp53tm1Brn Tg(Pdx1-cre/Esr1*)#Dam/J
038257 STOCK Krt13em1(cre/ERT2)Raja/J
036293 STOCK Lhx2tm1.1(cre/ERT2)Zjh/J
018418 STOCK Lrig1tm1.1(cre/ERT2)Rjc/J
039215 STOCK Robo3tm1.1(cre/ERT2)Ddg/J
032285 STOCK Slc34a1tm1(EGFP/cre/ERT2)Bhum/J
039562 STOCK Smr2tm1(cre)Ddg/J
039563 STOCK Sstr2tm1(cre/ERT2)Ddg/J
039022 STOCK Tacr1tm1.1(cre/ERT2)Ddg/J
039843 STOCK Tafa4em1.1(cre/ERT2)Ddg/J
040657 STOCK Tmprss2tm1.1(cre/ERT2)Ychen/J
039435 STOCK Tnctm1.1(cre/ERT2)Gcr/J
010911 STOCK Wt1tm1(EGFP/cre)Wtp/J
010912 STOCK Wt1tm2(cre/ERT2)Wtp/J
039156 STOCK Zic5tm1.1(icre/ERT2)Akg/J
020074 STOCK Tg(Dbh-cre/ERT2)198.1Hroh/J
039158 STOCK Tg(Dclk1-cre/ERT2)2Tcw/J
036851 STOCK Tg(KLKB1-cre/ERT2)1Pcn/CmoJ
005107 STOCK Tg(KRT14-cre/ERT)20Efu/J
028364 STOCK Tg(Neurod1-cre)1Able/J
012586 STOCK Tg(Slc1a3-cre/ERT)1Nat/J
018829 STOCK Tg(Sox9-cre/ERT2)1Msan/J
012708 STOCK Tg(Thy1-cre/ERT2,-EYFP)HGfng/PyngJ
115 strains use this protocol