Stock Number: 008183 Strain Name: B6.129S4(Cg)-Trp53tm2.1Tyj/J

Allele: Trp53tm2.1Tyj Protocol Name: Trp53tm4Tyj, tm2.1Tyj Method: Standard PCR Version: 1.1 Created: 12-September-2008 Updated: 24-September-2009

Expected Results: HET = 297 bp and 350 bp MUT = 350 bp WT = 297 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 oIMR8275 - AGC CTG CCT AGC TTC CTC AGG - Forward oIMR8276 - CTT GGA GAC ATA GCC ACA CTG - Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 10 X PCR Buffer II (ABI) 0.80 0.67 X 0.80 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR8275 0.80 1.33 uM 0.80 20 uM oIMR8276 0.80 1.33 uM 0.80 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 3 min - 2 94 30 sec - 3 64 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 008181 B6;129S4-Trp53tm4Tyj/J 008183 B6.129S4(Cg)-Trp53tm2.1Tyj/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.