Stock Number: 008250 Strain Name: STOCK Tg(Ins2-rtTA)2Efr/J

Allele: Generic tTA Protocol Name: Tg(tTA) Method: QPCR Version: 3.1 Created: 23-July -2008 Updated: 24-September-2009

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1544 - CAC GTG GGC TCC AGC ATT - Internal Positive Control Forward oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - Internal Positive Control Reverse oIMR4334 - AGC TCC ACT TAG ACG GCG AG - Transgene oIMR4335 - ATC GAA ATC GTC TAG CGC GT - Transgene TmoIMR0071 6-FAM GCA TGC GCC ATC GCC ACG TAMRA Transgene TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 Probe

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X Universal Master Mix 10.00 0.80 X 10.00 40 uM oIMR4334 0.25 0.40 uM 0.25 40 uM oIMR4335 0.25 0.40 uM 0.25 40 uM oIMR1544 0.25 0.40 uM 0.25 40 uM oIMR3580 0.25 0.40 uM 0.25 5 uM TmoIMR0071 0.75 0.15 uM 0.75 5 uM TmoIMR0107 0.75 0.15 uM 0.75 ddH2O 2.50 - 2.50 DNA 10.00 - 10.00

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 002618 B6.Cg-Tg(MMTVtTA)1Mam/J 003170 FVB.Cg-Tg(Myh6-tTA)6Smbf/J 003271 STOCK Tg(tTAhCMV)3Bjd/J 003272 STOCK Tg(tTALap)5Bjd/J 003273 STOCK Tg(rtTAhCMV)4Bjd/J 004127 FVB-Tg(Nes-rtTA)306Rvs/J 004937 NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ 005734 NOD/Lt-Tg(Ins2-rtTA)1Ach/AchJ 006235 B6.Cg-Tg(SFTPC-rtTA)5Jaw/J 006875 FVB/N-Tg(Tagln-rtTA)E1Jwst/J 006999 STOCK Dbttm1Geh Tg(tTALap)5Bjd Tg(tetO-DBT)A1Geh/J 007004 B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ 007678 B6;SJL-Tg(KRT14-rtTA)208Jek/J 008168 STOCK Tg(tetO-DTA)1Gfi/J 008250 STOCK Tg(Ins2-rtTA)2Efr/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.