Stock Number: 008008 Strain Name: SJL.129-Cd44tm1Ugu/J

Allele: Cd44tm1Ugu Protocol Name: Cd44tm1Ugu Method: Separated PCR Version: 1.1 Created: 08-February -2008 Updated: 24-September-2009

Expected Results: Mutant = 1300 bp Heterozygote = 1300 bp and 1500 bp Wild type = 1500 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 oIMR7826 - GAC CCA TTC GAG TCT CCT CTG GGG - Common oIMR7827 - ATG ATG ATG ATG GAG GAC AGA TAG - Wild type oIMR7828 - AAC GCT ATG TCC TGA TAG CGG T - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.20 - 4.20 10 X AB PCR BufferII 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 oIMR7826 0.60 1.00 uM 0.60 20 uM oIMR7828 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 5 min - 2 94 30 sec - 3 61.8 1 min - 4 72 1 min 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 4.20 - 4.20 10 X AB PCR BufferII 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 oIMR7826 0.60 1.00 uM 0.60 20 uM oIMR7827 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 008008 SJL.129-Cd44tm1Ugu/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.

Cycling  None found.