Stock Number: 006335 Strain Name: B6.129-Mgat5tm1Jwd/J

Allele: Mgat5tm1Jwd Protocol Name: Mgat5tm1Jwd Method: Standard PCR Version: 1.1 Created: 09-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 164 bp Heterozygote = 164 bp and 318 bp Wild type = 318 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 oIMR8068 - GCA AGG AAA CCA GCT GAC TC - Common oIMR8069 - TCC TCT GCC AGT GCC TTA AT - Wild type Reverse oIMR9012 - GTT TTC CCA GTC ACG ACG TT - 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 oIMR8068 0.60 1.00 uM 0.60 20 uM oIMR8069 0.60 1.00 uM 0.60 20 uM oIMR9012 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 64 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 006335 B6.129-Mgat5tm1Jwd/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.