Stock Number: 004301 Strain Name: 129-Trp53tm1Holl/J

Allele: Trp53tm1Holl Protocol Name: Trp53tm1Holl Method: Standard PCR Version: 1.1 Created: 02-May -2003 Updated: 24-September-2009

Expected Results: Mutant = 172 bp Heterozygote = 146 bp and 172 bp Wild type = 146 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1732 - GCC TCA TCT TGG GCC TGT GTT ATC TCC - Mutant oIMR1733 - GGC CAG TGT GCA GGG TGG CAA GTG GCT C - Mutant oIMR1734 - ACT CCA TGG CCC CTG TCA TC - Wild type oIMR1735 - AGG TCA CAC GAA AGA CAA CT - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 10 X PCR Buffer II (ABI) 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 oIMR1732 0.30 0.50 uM 0.30 20 uM oIMR1733 0.30 0.50 uM 0.30 20 uM oIMR1734 0.60 1.00 uM 0.60 20 uM oIMR1735 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 1.5 min - 2 94 30 sec - 3 66 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 004301 129-Trp53tm1Holl/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.