Stock Number: 003378 Strain Name: B6C3-Tg(APP695)3Dbo Tg(PSEN1)5Dbo/J

Allele: Generic Psen Protocol Name: Tg(PSEN1) Method: Melt Curve Analysis Version: 2.1 Created: 19-September-2008 Updated: 24-September-2009

Expected Results: Transgene Tm = 85.5°C +/- 1.0°C Internal positive control Tm = 79°C+/- 1.0°C

Gel Image

Image Image file View Full Size

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1644 - AAT AGA GAA CGG CAG GAG CA - Transgene oIMR1645 - GCC ATG AGG GCA CTA ATC AT - Transgene 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 1.20 - 1.20 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR1644 0.80 1.33 uM 0.80 20 uM oIMR1645 0.80 1.33 uM 0.80 20 uM oIMR7338 0.60 1.00 uM 0.60 20 uM oIMR7339 0.60 1.00 uM 0.60 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 15 sec - 3 62 1 min - 4 72 30 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 003378 B6C3-Tg(APP695)3Dbo Tg(PSEN1)5Dbo/J 004462 B6C3-Tg(APPswe,PSEN1dE9)85Dbo/J 005864 B6.Cg-Tg(APPswe,PSEN1dE9)85Dbo/J 005866 B6.Cg-Tg(APP695)3Dbo Tg(PSEN1dE9)S9Dbo/J 006554 B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/J 354462 B6C3-Tg(APPswe,PSEN1dE9)85Dbo 374139 Tg(APPswe,PSEN1dE9)85Dbo x C3H Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.