Stock Number: 003651 Strain Name: C57BL/6-Tg(Zp3-cre)93Knw/J

Allele: Generic Cre Protocol Name: Generic Cre Quantitative PCR Method: QPCR Version: 5.0 Created: 05-December -2011 Updated: 05-December -2011

Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/Quencher combination.  The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 13593 6-FAM AAA CAT GCT TCA TCG TCG GTC CGG Black Hole Quencher 1 Tg Probe oIMR1084 - GCG GTC TGG CAG TAA AAA CTA TC - Transgene Forward oIMR1085 - GTG AAA CAG CAT TGC TGT CAC TT - Transgene Reverse oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 IC Probe

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X PerfeCTa qPCR SuperMix 6.25 1.00 X 6.25 20 uM oIMR1084 0.13 0.21 uM 0.13 20 uM oIMR1085 0.13 0.21 uM 0.13 40 uM oIMR1544 0.13 0.42 uM 0.13 40 uM oIMR3580 0.13 0.42 uM 0.13 5 uM TmoIMR0107 0.38 0.15 uM 0.38 DNA 5.00 - 5.00 ddH2O 0.00 - 0.00 5 uM 13593 0.38 0.15 uM 0.38

Cycling  Step # Temp °C Time Note 1 50 2 min - 2 95 10 min - 3 95 15 sec - 4 60 1 min repeat steps 3-4 for 40 cycles

This genotyping protocol is used for the following strains:   Stock Number Strain Name 003314 FVB/N-Tg(EIIa-cre)C5379Lmgd/J 003328 129S/Sv-Tg(Prm-cre)58Og/J 003465 BALB/c-Tg(CMV-cre)1Cgn/J 003553 STOCK Tg(MMTV-cre)4Mam/J 003573 B6.Cg-Tg(Ins2-cre)25Mgn/J 003574 B6.Cg-Tg(Alb-cre)21Mgn/J 003651 C57BL/6-Tg(Zp3-cre)93Knw/J 003724 B6.FVB-Tg(EIIa-cre)C5379Lmgd/J 003771 B6.Cg-Tg(Nes-cre)1Kln/J 003802 B6.Cg-Tg(Lck-cre)548Jxm/J 004146 B6.129-Tg(Pcp2-cre)2Mpin/J 004426 B6;SJL-Tg(Cga-cre)3Sac/J 004746 STOCK Tg(Tagln-cre)1Her/J 004782 STOCK Tg(KRT14-cre)1Amc/J 005105 STOCK Tg(Chx10-EGFP/cre,-ALPP)2Clc/J 005107 STOCK Tg(KRT14-cre/ERT)20Efu/J 005359 B6.Cg-Tg(Camk2a-cre)T29-1Stl/J 005650 STOCK Tg(Myh6-cre/Esr1*)1Jmk/J 005657 B6.FVB(129)-Tg(Myh6-cre/Esr1*)1Jmk/J 006054 B6.C-Tg(CMV-cre)1Cgn/J 006137 B6.Cg-Tg(Cdh5-cre)7Mlia/J 006230 B6.Cg-Cebpatm1Dgt Tg(Mx1-cre)1Cgn/J 006410 B6;129S6-Chattm2(cre)Lowl/J 006774 FVB-Tg(Col2a1-cre/ERT)KA3Smac/J 006954 FVB-Tg(Ddx4-cre)1Dcas/J 007001 B6;129S-Tg(UBC-cre/ERT2)1Ejb/J 008085 B6.Cg-Tg(UBC-cre/ERT2)1Ejb/J 008454 B6.Cg-Tg(Sox2-cre)1Amc/J 008533 B6;FVB-Tg(Cspg4-cre)1Akik/J 008766 C57BL/6-Tg(Cd8a-cre)1Itan/J 008851 STOCK Tg(Wnt1-cre/ERT)1Alj/J 010527 B6;129-Gt(ROSA)26Sortm1(DTA)Mrc/J 012686 C57BL/6N-Tg(Ppp1r2-cre)4127Nkza/J 012899 STOCK Agrptm1(cre)Lowl/J 016261 C57BL/6-Tg(Nes-cre/ERT2)KEisc/J 017526 B6.129-Nos1tm1(cre)Mgmj/J 1000075 STOCK Tg(Myh6-cre/Esr1*)1Jmk/J (also maybe) Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.