Stock Number: 008841 Strain Name: B6.129S2-Ccrn4ltm1Bjc/J

Allele: Ccrn4ltm1Bjc Protocol Name: Ccrn41tm1Bjc Method: Standard PCR Version: 1.1 Created: 01-May -2009 Updated: 14-December -2009

Expected Results: Mutant = 550 bp Heterozygote = 410 bp and 550 bp Wild type = 410 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 oIMR9687 - CTC CAG AAC CTG CAG AAC ATC - Wild type oIMR9688 - AAC ATC CCT GGT TAA GAC GCT - Common oIMR9689 - GCC TTC TAT CGC CTT CTT GAC - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.16 - 3.16 10 X AB PCR BufferII 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 oIMR9687 0.60 1.00 uM 0.60 20 uM oIMR9688 0.60 1.00 uM 0.60 20 uM oIMR9689 0.80 1.33 uM 0.80 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 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 008841 B6.129S2-Ccrn4ltm1Bjc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.