Stock Number: 007049 Strain Name: B6.Cg-Tg(tetO-APPSwInd)885Dbo/J

Allele: Generic App Protocol Name: Tg(APP) Method: Standard PCR Version: 2.1 Created: 28-July -2008 Updated: 24-September-2009

Expected Results: Transgene = 377 bp Internal positive control = 324 bp

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

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 oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.12 - 3.12 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR3610 0.60 1.00 uM 0.60 20 uM oIMR3611 0.60 1.00 uM 0.60 20 oIMR8744 0.30 0.50 0.30 20 uM oIMR8745 0.30 0.50 uM 0.30 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 52 1 min - 4 72 1 min 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 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 Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.