Stock Number: 002197 Strain Name: C57BL/6-Col1a1Mov13/J

Allele: Col1a1Mov13 Protocol Name: Col1a1Mov13 Method: Standard PCR Version: 1.1 Created: 01-February -2002 Updated: 24-September-2009

Expected Results: Mutant = 283 bp Heterozygote = 135 bp and 283 bp Wild type = 135 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 oIMR0145 - TCA GCT TTG TGG ACC TCC GG - Common oIMR0146 - GAC CCC TCT ATA CAG AAC GC - Wild type oIMR0147 - CTT CTG CTC CCC GAG CTC AA - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.68 - 3.68 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.48 0.10 mM 0.48 20 uM oIMR0145 0.72 1.20 uM 0.72 20 uM oIMR0146 0.39 0.65 uM 0.39 20 uM oIMR0147 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 0.07 0.03 U/ul 0.07 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 56 30 sec - 4 72 30 sec 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 002197 C57BL/6-Col1a1Mov13/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.