Stock Number: 006607 Strain Name: B6.129-Pctptm1Bor/J

Allele: Pctptm1Bor Protocol Name: Pctptm1Bor Method: Standard PCR Version: 1.1 Created: 30-June -2008 Updated: 24-September-2009

Expected Results: Mutant = 184 bp Heterozygote = 184 bp and 369 bp Wild type = 369 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 oIMR6464 - GTT TGG GGT GAC AAG CAA GT - - oIMR6465 - GGG CAA AGA GCA CAT CAA AT - Common oIMR7415 - GCC AGA GGC CAC TTG TGT AG - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR6464 0.60 1.00 uM 0.60 20 uM oIMR6465 0.60 1.00 uM 0.60 20 uM oIMR7415 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 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 006607 B6.129-Pctptm1Bor/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.