Stock Number: 002829 Strain Name: B6.129P2-Plaurtm1Jld/J

Allele: Plaurtm1Jld Protocol Name: Plaurtm1Jld Method: Standard PCR Version: 3.1 Created: 23-October -2008 Updated: 24-September-2009

Expected Results: Mutant = 200bp Heterozygote = 398bp and 200bp Wild type = 398bp

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 oIMR0491 - TAT TAC CAG TGA ATC TTT GTC AGC AGT TCC C - Mutant oIMR3172 - AGA GCT CCG GGT TCT CTC TC - Common oIMR3173 - GGG AGG AAG GAA CTC CAC TC - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.56 - 3.56 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 oIMR3172 0.80 1.33 uM 0.80 20 uM oIMR3173 0.60 1.00 uM 0.60 20 uM oIMR0491 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 68 45 sec - 4 72 45 sec 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 002829 B6.129P2-Plaurtm1Jld/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.