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

Allele: Bace1tm1Pcw Protocol Name: Bace1tm1Pcw Method: Standard PCR Version: 1.1 Created: 11-May -2004 Updated: 24-September-2009

Expected Results: Mutant = 157 bp Heterozygote = 272 bp and 157 bp Wild type = 272 bp

Gel Image

Image Image file View Full Size

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

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 3.36 - 3.36 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 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 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 58 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 8 72 2 min - 9 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.