Stock Number: 008597 Strain Name: B6.129-Ppargc1atm1Brsp/J

Allele: Ppargc1atm1Brsp Protocol Name: Ppargc1asuptm1Brsp/sup Method: Standard PCR Version: 2.0 Created: 20-July -2010 Updated: 20-July -2010

Expected Results: Mutant = 400 bp Heterozygote = 400 bp and 648 bp Wild type = 648 bp

Gel Image

no data found

Gel Information: Separate on gel electrophoresis on 1.5% agarose gel

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type 8039 - CCA GTT TCT TCA TTG GTG TG - Wild type Forward 8040 - ACC TGT CTT TGC CTA TGA TTC - Wild type Reverse 8041 - TCC AGT AGG CAG AGA TTT ATG AC - Mutant Forward 8042 - CCA ACT GTC TAT AAT TCC AGT TC - Mutant Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 DNA 2.00 - 2.00 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 10 X AB PCR BufferII 1.20 1.00 X 1.20 ddH2O 2.76 - 2.76 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM 8039 0.60 1.00 uM 0.60 20 uM 8040 0.60 1.00 uM 0.60 20 uM 8042 0.60 1.00 uM 0.60 20 uM 8041 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30sec - 3 64 1 min - 4 72 1 min repeat steps 2, 3 and 4 - 35 times 5 72 2 min - 6 10 - Hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 008597 B6.129-Ppargc1atm1Brsp/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.