Stock Number: 003201 Strain Name: 129/Sv-Csktm1Sor/J

Allele: Csktm1Sor Protocol Name: Csktm1Sor Method: Standard PCR Version: 1.1 Created: 20-August -2008 Updated: 24-September-2009

Expected Results: Mutant = ~470 bp Heterozygote = 225 bp and ~470 bp Wild type = 225 bp

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1867 - CGC AGT CTA CGA GGT GAT GA - Wild type oIMR1868 - GGG CTC AGT TCA AGT TCA GG - Common oIMR6218 - CCT TCT ATC GCC TTC TTG ACG - Mutant

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 oIMR1868 0.60 1.00 uM 0.60 20 uM oIMR1867 0.60 1.00 uM 0.60 20 uM oIMR6218 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 60 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 003201 129/Sv-Csktm1Sor/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.