Stock Number: 008333 Strain Name: B6;129P2-Dldtm1Ptl/J

Allele: Dldtm1Ptl Protocol Name: Dldtm1Ptl Method: Standard PCR Version: 1.1 Created: 14-July -2008 Updated: 24-September-2009

Expected Results: Mutant = 232 bp Heterozygote = 232 bp and 340 bp Wild type = 340 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 oIMR6218 - CCT TCT ATC GCC TTC TTG ACG - Mutant oIMR8434 - CAT GGC TCC TTT CAG CTG TT - Wild type Forward oIMR8435 - ACA ATA TAC CCG CCT CAC CA - Common oIMR8436 - TCG CCT TCT TGA CGA GTT CT - 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 oIMR8434 0.60 1.00 uM 0.60 20 uM oIMR8435 0.60 1.00 uM 0.60 20 uM oIMR8436 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 61.5 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 008333 B6;129P2-Dldtm1Ptl/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.