Stock Number: 005864 Strain Name: B6.Cg-Tg(APPswe,PSEN1dE9)85Dbo/J

Allele: Generic App Protocol Name: Generic Tg(APP) Melt Curve Analysis Method: Melt Curve Analysis Version: 3.1 Created: 18-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

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR3610 - AGG ACT GAC CAC TCG ACC AG - Transgene oIMR3611 - CGG GGG TCT AGT TCT GCA T - 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 1.60 - 1.60 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR7338 0.60 1.00 uM 0.60 20 uM oIMR7339 0.60 1.00 uM 0.60 20 uM oIMR3610 0.60 1.00 uM 0.60 20 uM oIMR3611 0.60 1.00 uM 0.60 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 50.5 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 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 006004 B6C3-Tg(tetO-APPSwInd)885Dbo/J 006005 B6.Cg-Tg(Prnp-App/APPswe)E1-2Dbo/J 006006 B6.Cg-Tg(Prnp-APP)A-2Dbo/J 006554 B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/J 007049 B6.Cg-Tg(tetO-APPSwInd)885Dbo/J 007051 B6.Cg-Tg(tetO-APPSwInd)102Dbo/J 007052 B6.Cg-Tg(tetO-APPSwInd)107Dbo/J 009126 B6.Cg-Nos2tm1Lau Tg(Thy1-APPSwDutIowa)BWevn/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.