Stock Number: 007182 Strain Name: B6.Cg-Tg(Prnp-ITM2B/APP695*42)A12Emcg/J

Allele: Tg(Prnp-ITM2B/APP695*42)A12Emcg Protocol Name: Tg(Prnp-ITM2B/APP695*42)A12Emcg MCA Method: Melt Curve Analysis Version: 1.0 Created: 04-August -2009 Updated: 24-September-2009

Expected Results: Tg/0 Area = 5 Tg/0 Tm = 80.2 +/- 1 Tg/02 Area = .5 Tg/02 Tm = 87.8 +/- 1 internal control Area = 10 internal control Tm = 87.8 +/- 1

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0647 - CAG ACA TCG AGG AAG TGA AGG - Internal Positive Control Forward oIMR0648 - TGC TGT GGT CAT CAG GTA GG - Internal Positive Control Reverse oIMR7042 - AAG GCT GGA ACC TAT TTG CC - Transgene Forward oIMR7044 - TAG TGG ATC CCT ACG CTA TG - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.00 - 2.00 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR7042 0.80 1.33 uM 0.80 20 uM oIMR7044 0.80 1.33 uM 0.80 20 uM oIMR0647 0.20 0.33 uM 0.20 20 uM oIMR0648 0.20 0.33 uM 0.20 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 65 1 min - 4 72 45 sec 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 007002 B6;C3-Tg(Prnp-ITM2B/APP695*42)A12Emcg/J 007182 B6.Cg-Tg(Prnp-ITM2B/APP695*42)A12Emcg/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.