Stock Number: 005701 Strain Name: STOCK Pdx1tm1Macd/J

Allele: Pdx1tm1Macd Protocol Name: Pdx1tm1Macd Method: Standard PCR Version: 1.1 Created: 17-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 660 bp Heterozygote = 520 bp and 660 bp Wild type = 520 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 oIMR4260 - ACC ATG AAC AGT GAG GAG CAG TAC - Wild type oIMR4261 - GCG GGT TTC AGA GGA ACT TGT - Wild type oIMR4262 - TAG AAG GGG AAA GCT GGC AAG - Mutant oIMR4263 - TCC AGA TCG AAA TCG TCT AGC G - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.72 - 2.72 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 oIMR4260 0.80 1.33 uM 0.80 20 uM oIMR4261 0.80 1.33 uM 0.80 20 uM oIMR4262 0.40 0.67 uM 0.40 20 uM oIMR4263 0.40 0.67 uM 0.40 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.10 0.04 U/ul 0.10 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 68 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 005701 STOCK Pdx1tm1Macd/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.