Stock Number: 006665 Strain Name: B6;129P2-Olfr151tm13(rI7)Mom/MomJ

Allele: Olfr151tm13(Olr226)Mom Protocol Name: Olfr151tm13(olr226)Mom Method: Separated PCR Version: 1.1 Created: 01-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 500 bp Heterozygote = 309 bp and 500 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 mom0075 - GGA TCT GCC ATT GTC AGA CAT G - Mutant oIMR7149 - GCA GCT GTG GGC TTG TTC T - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.26 - 4.26 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 mom0008 0.60 1.00 uM 0.60 20 uM mom0075 0.30 0.50 uM 0.30 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 95 5 min - 2 95 1 min - 3 60 1 min - 4 72 2 min repeat steps 2-4 for 35 cycles 5 72 10 min - 6 10 - hold

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.26 - 4.26 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 oIMR7149 0.30 0.50 uM 0.30 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 006665 B6;129P2-Olfr151tm13(rI7)Mom/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 95 5 min - 2 95 1 min - 3 60 1 min - 4 72 2 min repeat steps 2-4 for 35 cycles 5 72 10 min - 6 10 - hold