Stock Number: 004584 Strain Name: B6.129-Ppargtm2Rev/J

Allele: Ppargtm2Rev Protocol Name: Ppargtm2Rev Method: Standard PCR Version: 1.1 Created: 05-October -2004 Updated: 24-September-2009

Expected Results: Mutant = 230 bp Heterozygote = ~200 bp and 230 bp Wild type = ~200 bp

Gel Image

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Gel Information: Separated by gel electrophoresis on a 3.0% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1934 - TGT AAT GGA AGG GCA AAA GG - - oIMR1935 - TGG CTT CCA GTG CAT AAG TT - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 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 oIMR1934 0.60 1.00 uM 0.60 20 uM oIMR1935 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 60 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 8 72 2 min - 9 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 004584 B6.129-Ppargtm2Rev/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.