Stock Number: 003322 Strain Name: B6.129S4-Soat1tm1Far/J

Allele: Soat1tm1Far Protocol Name: Soat tm1Far Method: Standard PCR Version: 1.1 Created: 12-February -2001 Updated: 24-September-2009

Expected Results: Mutant = 280 bp Heterozygote = 280 bp and ~700 bp Wild type = ~700bp

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 oIMR0740 - CAG CTA TGT ACA CAC ATA TAT TCA TG - Wild type oIMR0741 - GGA CTT TTC AAT GAG GTT GGT CAC - Wild type oIMR6916 - CTT GGG TGG AGA GGC TAT TC - Mutant Forward oIMR6917 - AGG TGA GAT GAC AGG AGA TC - 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 oIMR6916 0.30 0.50 uM 0.30 20 uM oIMR6917 0.30 0.50 uM 0.30 20 uM oIMR0740 0.60 1.00 uM 0.60 20 uM oIMR0741 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 1.5 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 002896 B6;129S4-Soat1tm1Far/J 003322 B6.129S4-Soat1tm1Far/J 007147 B6.129S4-Soat1tm1Far/Pgn Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.