Stock Number: 007084 Strain Name: B6.FVB(Cg)-Mmp9tm1Tvu/J

Allele: Mmp9tm1Tvu Protocol Name: Mmp9 tm1Tvu Method: Standard PCR Version: 1.1 Created: 04-April -2003 Updated: 10-November -2009

Expected Results: Mutant = 172 bp Heterozygote = 172 bp and 277 bp Wild type = 277bp

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 oIMR0158 - CTG AAT GAA CTG CAG GAC GA - Mutant oIMR0159 - ATA CTT TCT CGG CAG GAG CA - Mutant oIMR1760 - GTG GGA CCA TCA TAA CAT CAC A - Wild type oIMR1761 - CTC GCG GCA AGT CTT CAG AGT A - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.76 - 2.76 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 oIMR1760 0.60 1.00 uM 0.60 20 uM oIMR1761 0.60 1.00 uM 0.60 20 uM oIMR0158 0.60 1.00 uM 0.60 20 uM oIMR0159 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 66 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 004104 FVB.Cg-Mmp9tm1Tvu/J 007084 B6.FVB(Cg)-Mmp9tm1Tvu/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.