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Stock No: 029211
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 |
|---|---|
| 037511 | B6(129S4)-Pou2f3tm1.1(cre/ERT2)Imt/J |
| 038514 | B6(129S6)-Tg(Slc6a5-cre/ERT2)#Uze/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 |
| 038495 | B6.129S6(Cg)-Gli1tm3(cre/ERT2)Alj/RtigJ |
| 038129 | B6.Cg-Ascl3tm2.1(EGFP/icre/ERT2)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 |
| 038834 | B6.Cg-Sdk1tm1.1(cre/ERT2)Jrs/KrishJ |
| 038840 | B6.Cg-Sdk2tm1.1(cre/ERT2)Jrs/KrishJ |
| 029567 | B6.Cg-Tg(Col1a2-cre/ERT,-ALPP)7Cpd/2J |
| 016237 | B6.Cg-Tg(Col1a2-cre/ERT,-ALPP)7Cpd/J |
| 008538 | B6.Cg-Tg(Cspg4-cre/Esr1*)BAkik/J |
| 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/AkinJ |
| 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 |
| 036292 | B6;129-Gt(ROSA)26Sortm1(BEST1-cre/ERT2)Vold/J |
| 036300 | B6;129S4-Cux1tm1(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 |
| 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 |
| 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 |
| 038831 | C57BL/6-Efnb2em1(cre/ERT2)Kloh/J |
| 038640 | C57BL/6-Rtkn2em1(cre/ERT2)Jich/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 |
| 039217 | C57BL/6J-Dpp4em1(cre/ERT2)Jkpns/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 |
| 036942 | STOCK Agertm2.1(cre/ERT2)Blh/2J |
| 036343 | STOCK Apoc4em1(cre/ERT2)Hzhu/J |
| 036246 | STOCK Cyp1a2em1(cre/ERT2)Hzhu/J |
| 038582 | STOCK Fbxo2tm1.1(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 |
| 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 |
| 032285 | STOCK Slc34a1tm1(EGFP/cre/ERT2)Bhum/J |
| 039022 | STOCK Tacr1tm1.1(cre/ERT2)Ddg/J |
| 010911 | STOCK Wt1tm1(EGFP/cre)Wtp/J |
| 010912 | STOCK Wt1tm2(cre/ERT2)Wtp/J |
| 025736 | STOCK Tg(Cela1-cre/ERT)1Lgdn/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 |
| 81 strains use this protocol | |