Stock Number: 004714 Strain Name: B6.129-Bace1tm1Pcw/J

Allele: Bace1tm1Pcw Protocol Name: Bacetm1Pcw Method: Melt Curve Analysis Version: 1.0 Created: 26-May -2009 Updated: 24-September-2009

Expected Results: Mutant Tm = 87.5°C +/- 1.0°C Wild type Tm = 85.2°C+/- 1.0°C

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR3169 - AGG CAG CTT TGT GGA GAT GGT G - Wild type oIMR3170 - CGG GAA ATG GAA AGG CTA CTC C - Common oIMR8162 - TGG ATG TGG AAT GTG TGC GAG - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.20 - 2.20 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR3169 0.60 1.00 uM 0.60 20 uM oIMR3170 0.60 1.00 uM 0.60 20 uM oIMR8162 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 69 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 004714 B6.129-Bace1tm1Pcw/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.