Stock Number: 010673 Strain Name: B6;129-Runx1tm3.1Spe/J

Allele: Runx1tm3.1Spe Protocol Name: Runx1tm3.1Spe Method: Standard PCR Version: 2.0 Created: 16-June -2010 Updated: 16-June -2010

Expected Results: Mutant = ~275 bp Heterozygote = ~275 bp and 203 bp Wild type = 203 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 9297 - CCC ACT GTG TGC ATT CCA GAT TGG - Forward 9298 - GAC GGT GAT GGT CAG AGT GAA GC - Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.92 - 4.92 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 DNA 2.00 - 2.00 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM 9297 0.60 1.00 uM 0.60 20 uM 9298 0.60 1.00 uM 0.60 10 X KAPA Rxn Buffer 1.20 1.00 X 1.20 5 U/ul KAPATaq DNA Polymerase 0.06 0.03 U/ul 0.06

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 65 30 sec - 4 72 1 min - 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 010673 B6;129-Runx1tm3.1Spe/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.