Stock Number: 006492 Strain Name: B6.Cg-Pdgfratm8Sor/EiJ

Allele: Pdgfratm8Sor Protocol Name: Pdgfratm8Sor Method: Standard PCR Version: 1.1 Created: 30-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 242 bp Heterozygote = 242 bp and 451 bp Wild type = 451 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 oIMR7801 - CCC TTG TGG TCA TGC CAA AC - Wild type Forward oIMR7802 - GCT TTT GCC TCC ATT ACA CTG G - Wild type Reverse oIMR7919 - ACG AAG TTA TTA GGT CCC TCG AC - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 4.56 - 4.56 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 oIMR7801 0.20 0.33 uM 0.20 20 uM oIMR7802 0.20 0.33 uM 0.20 20 uM oIMR7919 0.20 0.33 uM 0.20 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 65 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 006492 B6.Cg-Pdgfratm8Sor/EiJ Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.