Stock Number: 006866 Strain Name: STOCK Tg(Ins1-DsRed*T4)32Hara/J

Allele: Generic DsRed Protocol Name: Generic DsRed Method: Melt Curve Analysis Version: 5.0 Created: 17-June -2010 Updated: 17-June -2010

Expected Results: Transgene Tm = 87.9°C +/- 1.0°C Internal positive control Tm = 82.7°C+/- 1.0°C

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6787 - CCC ATG GTC TTC TTC TGC AT - Transgene Forward oIMR6788 - AAG GTG TAC GTG AAG CAC CC - Transgene Reverse oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 1.60 - 1.60 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR6787 0.80 1.33 uM 0.80 20 uM oIMR6788 0.80 1.33 uM 0.80 20 uM oIMR7338 0.40 0.67 uM 0.40 20 uM oIMR7339 0.40 0.67 uM 0.40 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 se - 3 68 1 min - 4 72 45 se repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 005328 NOD/ShiLt-Tg(Cd4-DsRed)4Lt/J 005441 STOCK Tg(CAG-DsRed*MST)1Nagy/J 006051 B6.Cg-Tg(CAG-DsRed*MST)1Nagy/J 006055 B6.Cg-Tg(CAG-Bgeo,-DsRed*MST)1Nagy/J 006866 STOCK Tg(Ins1-DsRed*T4)32Hara/J 006872 B6.Cg-Tg(Ins1-DsRed*T4)32Hara/J 008241 STOCK Tg(Cspg4-DsRed.T1)1Akik/J 008605 B6;C3-Tg(CAG-DsRed,-EGFP)5Gae/J 008705 B6.Cg-Tg(CAG-DsRed,-EGFP)5Gae/J 009655 C57BL/6J-Tg(Dcx-DsRed)14Qlu/J 017620 NOD.Cg-Prkdcscid Tg(CAG-DsRed*MST)1Nagy/KupwJ 018410 B6.Cg-Tg(Pnkd*A7V*A9V,-DsRed)671Ljp/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.