Stock Number: 007027 Strain Name: C57BL/6-Tg(Thy1-APPSwDutIowa)BWevn/J

Allele: Generic App Protocol Name: Tg(APP) Method: Separated QPCR Version: 2.0 Created: 02-November -2009 Updated: 02-November -2009

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 5 uM TmoIMR0076 0.38 0.15 uM 0.38 ddH2O 0.64 - 0.64 DNA 5.00 - 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

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 oIMR1544 0.13 0.42 uM 0.13 40 uM oIMR3580 0.13 0.42 uM 0.13 5 uM TmoIMR0107 0.38 0.15 uM 0.38 ddH2O 0.64 - 0.64 DNA (crude prep diluted 1:100 in water) 5.00 - 5.00 This genotyping protocol is used for the following strains:   Stock Number Strain Name 007027 C57BL/6-Tg(Thy1-APPSwDutIowa)BWevn/J 009126 B6.Cg-Nos2tm1Lau Tg(Thy1-APPSwDutIowa)BWevn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  Step # Temp °C Time Note 5 50 2 min - 6 95 10 min - 7 95 15 sec - 8 60 1 min repeat steps 7- for 40 cycles