Stock Number: 002659 Strain Name: FVB/N-Tg(Trp53R172H)8512Jmr/J

Allele: Tg(Trp53R172H)8512Jmr Protocol Name: Tg(Trp53R172_) Method: Standard PCR Version: 1.1 Created: 01-September-2000 Updated: 24-September-2009

Expected Results: Transgene = 1100 bp Internal positive control = ~500 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 oIMR0415 - GCC TGA AAA TGT CTC CTG GCT CAG AGG G - Transgene oIMR0416 - GAT GTG CTC CAG GCT AAA GTT - - oIMR0417 - AGA AAC GGA ATG TTG TGG AGT - - oIMR7951 - CCG TCG ACG GCC ACA GTG AAG ACC TCC GGC CAG - Forward

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.00 - 3.00 10 X PCR Buffer II (ABI) 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 oIMR7951 0.60 1.00 uM 0.60 20 uM oIMR0415 0.60 1.00 uM 0.60 20 uM oIMR0416 0.60 1.00 uM 0.60 20 uM oIMR0417 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 3 min - 2 94 20 sec - 3 64 30 sec* *-0.5 C per cycle 4 72 35 sec repeat steps 2-4 for 12 cycles 5 94 20 sec - 6 58 30 sec - 7 72 35 sec repeat steps 5-7 for 25 cycles 8 72 2 min - 9 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 002659 FVB/N-Tg(Trp53R172H)8512Jmr/J 002660 FVB/N-Tg(Trp53R172L)4491Jmr/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.