Stock Number: 006675 Strain Name: B6;129P2-Olfr151tm25Mom/MomJ

Allele: Olfr151tm25Mom Protocol Name: Olfr151tm25Mom Method: Standard PCR Version: 1.1 Created: 01-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 250 bp Heterozygote = 250 bp and 309 bp Wild type = 309 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 mom0140 - TGG AGC CCG TCA GTA TCG GC - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.60 - 3.60 10 X PCR Buffer II (ABI) 0.96 0.80 X 0.96 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM mom0140 0.60 1.00 uM 0.60 20 uM mom0008 0.60 1.00 uM 0.60 20 uM mom0135 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 58.4 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 006675 B6;129P2-Olfr151tm25Mom/MomJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.