Stock Number: 005078 Strain Name: NOD.Cg-Il1r1tm1Roml/HetJ

Allele: Il1r1tm1Roml Protocol Name: Il1r1tm1Roml Method: Standard PCR Version: 1.1 Created: 13-March -2001 Updated: 24-September-2009

Expected Results: Mutant = 172 bp Heterozygote = 172 bp and 350 bp Wild type = 350 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 oIMR0158 - CTG AAT GAA CTG CAG GAC GA - Mutant oIMR0159 - ATA CTT TCT CGG CAG GAG CA - Mutant oIMR0779 - CCA CAT ATT CTC CAT CAT CTC TGC TGG TA - Wild type oIMR0780 - TTT CGA ATC TCA GTT GTC AAG TGT GTC CC - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.76 - 2.76 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 oIMR0158 0.60 1.00 uM 0.60 20 uM oIMR0159 0.60 1.00 uM 0.60 20 uM oIMR0779 0.60 1.00 uM 0.60 20 uM oIMR0780 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 63 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 003018 B6;129S1-Il1r1tm1Roml/J 005078 NOD.Cg-Il1r1tm1Roml/HetJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.