Stock Number: 008087 Strain Name: B6.129S1-Bchetm1Loc/J

Allele: Bchetm1Loc Protocol Name: Bchetm1Loc Method: Standard PCR Version: 1.1 Created: 08-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 240 bp Heterozygote = 240 bp and 337 bp Wild type = 337 bp

Gel Image

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR7415 - GCC AGA GGC CAC TTG TGT AG - Mutant oIMR8093 - CAT GGA AAA GAG GAG GAT GG - Common oIMR8094 - TTC CAC TTG GAA GAG AAA CAC A - Wild type Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.60 - 3.60 10 X PCR Buffer II (ABI) 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8093 0.60 1.00 uM 0.60 20 uM oIMR8094 0.60 1.00 uM 0.60 20 uM oIMR7415 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 5 min - 2 94 30 sec - 3 61 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 008077 129S1/Sv-Bchetm1Loc/J 008087 B6.129S1-Bchetm1Loc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.