Stock Number: 007176 Strain Name: B6.Cg-Tg(Pax8-rtTA2S*M2)1Koes/J

Allele: Tg(Pax8-rtTA2S*M2)1Koes Protocol Name: Tg(Pax8-rtTA2S*M2)1Koes Method: QPCR Version: 2.1 Created: 23-October -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 oIMR7387 - CTG GAG AAC GCA CTG TAC GC - Transgene Forward oIMR7388 - CCA ATA CGC AGC CCA GTG T - Transgene Reverse TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 Probe TmoIMR0110 6-FAM CTG TCC GCC GTG GGC CAC TT TAMRA Transgene

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 oIMR7387 0.25 0.40 uM 0.25 40 uM oIMR7388 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 TmoIMR0110 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 007176 B6.Cg-Tg(Pax8-rtTA2S*M2)1Koes/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.