Stock Number: 005300 Strain Name: B6.129-Tg(APPSw)40Btla/J

Allele: Tg(APPSw)40Btla Protocol Name: Tg(APPSw)40Btla Method: Standard PCR Version: 1.1 Created: 16-June -2008 Updated: 05-November -2009

Expected Results: TG=84 bp WT=200 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 oIMR6938 - CTT CAC TCG TTC TCA TTC TCT TCC A - Transgene oIMR6939 - GCG TTT TTA TCC GCA TTT CGT TTT 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 2.76 - 2.76 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR6938 0.60 1.00 uM 0.60 20 uM oIMR6939 0.60 1.00 uM 0.60 20 oIMR8744 0.60 1.00 0.60 20 uM oIMR8745 0.60 1.00 uM 0.60 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 60 30 sec - 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 005300 B6.129-Tg(APPSw)40Btla/J 006406 B6.129S4-Tg(APPSwLon)96Btla/J 006409 129S1.129(Cg)-Tg(APPSw)40Btla/J 006472 D2.129(B6)-Tg(APPSw)40Btla/J 006555 A.129(B6)-Tg(APPSw)40Btla/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.