Stock Number: 005642 Strain Name: B6.Cg-Clutm1Jakh/J

Allele: Clutm1Jakh Protocol Name: Clutm1Jakh Method: Standard PCR Version: 1.1 Created: 18-July -2006 Updated: 24-September-2009

Expected Results: Mutant = 500 bp Heterozygote = 500 bp and 540 bp Wild type = 540 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 oIMR4020 - ACG ATG TGG AAG GAT GTG GAA GAT GAA CAT G - Common oIMR4021 - TGG TGA TGG GGC TCT AGT CAC CTC CCA CCT C - - oIMR4022 - CTG CTA AAG CGC ATG CTC CAG ACT GCC TTG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.66 - 3.66 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 oIMR4020 0.60 1.00 uM 0.60 20 uM oIMR4021 0.60 1.00 uM 0.60 20 uM oIMR4022 0.30 0.50 uM 0.30 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 70 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 005642 B6.Cg-Clutm1Jakh/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.