Stock Number: 008609 Strain Name: 129S1.129(Cg)-Tg(APPSw)40Btla/2J

Allele: Generic App Protocol Name: Tg(APP) Method: QPCR Version: 3.0 Created: 02-November -2009 Updated: 02-November -2009

Notes: Taqman qPCR protocols are run on an ABI 7500, 7700, 7900 or the Roche LightCycler 480. Use an appropriate instrument-specific Fuorophore/Quencher combination.  The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1544 - CAC GTG GGC TCC AGC ATT - - oIMR3580 - TCA CCA GTC ATT TCT GCC TTT G - - oIMR5260 - TGG GTT CAA ACA AAG GTG CAA - - oIMR5261 - GAT GAA GAT CAC TGT CGC TAT GAC - - TmoIMR0076 6-FAM CAT TGG ACT CAT GGT GGG CGG TG TAMRA - TmoIMR0107 JOE NHS Ester CCA ATG GTC GGG CAC TGC TCA A Black Hole Quencher 1 -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 2 X Universal Master Mix 6.25 1.00 X 6.25 40 uM oIMR5260 0.13 0.42 uM 0.13 40 uM oIMR5261 0.13 0.42 uM 0.13 40 uM oIMR1544 0.13 0.42 uM 0.13 40 uM oIMR3580 0.13 0.42 uM 0.13 5 uM TmoIMR0076 0.38 0.15 uM 0.38 5 uM TmoIMR0107 0.38 0.15 uM 0.38 DNA 5.00 0.00 5.00

Cycling  Step # Temp °C Time Note 1 50 2 min - 2 95 10 min - 3 95 15 sec - 4 60 1 min repeat steps 3-4 for 40 cycles

This genotyping protocol is used for the following strains:   Stock Number Strain Name 004661 B6.Cg-Tg(PDGFB-APPSwInd)20Lms/1J 005300 B6.129-Tg(APPSw)40Btla/J 005301 B6.129S2-Tg(APP)8.9Btla/J 006293 B6.Cg-Tg(PDGFB-APPSwInd)20Lms/2J 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 008609 129S1.129(Cg)-Tg(APPSw)40Btla/2J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.