Stock Number: 009120 Strain Name: B6.129P2-Axin2tm1Wbm/J

Allele: Axin2tm1Wbm Protocol Name: Axin2tm1Wbm Method: Standard PCR Version: 1.0 Created: 06-October -2010 Updated: 14-January -2011

Expected Results: Mutant = 400 bp Heterozygote = 400 bp and 493 bp Wild type = 493 bp

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 9391 - AAG CTG CGT CGG ATA CTT GAG A - Wild type Forward 9392 - AGT CCA TCT TCA TTC CGC CTA GC - Wild type Reverse 9393 - TGG TAA TGC TGC AGT GGC TTG - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 2.5 mM dNTP 0.96 0.20 mM 0.96 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 25 mM MgCl2 0.96 2.00 mM 0.96 DNA 2.00 - 2.00 20 uM 9393 0.60 1.00 uM 0.60 20 uM 9392 0.60 1.00 uM 0.60 10 X AB PCR BufferII 1.20 1.00 X 1.20 20 uM 9391 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 67 30 sec - 4 72 30 sec - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 009120 B6.129P2-Axin2tm1Wbm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.