Stock Number: 006648 Strain Name: B6;129P2-Olfr151tm22Mom/MomJ

Allele: Olfr151tm22Mom Protocol Name: Olfr151tm22Mom Method: Separated PCR Version: 1.1 Created: 01-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 250 bp Heterozygote = 250 bp and 300 bp Wild type = 300 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 mom0008 - ATG GAG AGC AAT GAT TCC TG - Common mom0135 - GCA GCT GTG GGC TTG TTC TAT GGA TCC - Wild type mom0139 - GAG AAG CGC GAT CAC ATG GTC CT - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 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 mom0008 0.60 1.00 uM 0.60 20 uM mom0139 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 2 min - 2 94 30 sec - 3 60 45 sec - 4 72 45 sec repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 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 mom0135 0.60 1.00 uM 0.60 20 uM mom0008 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 This genotyping protocol is used for the following strains:   Stock Number Strain Name 006648 B6;129P2-Olfr151tm22Mom/MomJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  Step # Temp °C Time Note 1 94 2 min - 2 94 30 sec - 3 60 45 sec - 4 72 45 sec repeat steps 2-4 for 35 cycles 5 72 2 min - 6 10 - hold