Stock Number: 006618 Strain Name: C57BL/6-Tg(tetO-COX8A/EYFP)1Ksn/J

Allele: Tg(tetO-COX8A/EYFP)1Ksn Protocol Name: Tg(tetO-COX8A/EYFP)1Ksn Method: Standard PCR Version: 2.0 Created: 09-January -2010 Updated: 09-January -2010

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 338 bp Internal positive control = 200 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 oIMR6435 - GCT GAC CCT GAA GTT CAT CTG C - Transgene oIMR6436 - CAT GAT ATA GAC GTT GTG GCT GTT G - Transgene oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.54 - 3.54 10 X AB PCR BufferII 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 oIMR6435 0.60 1.00 uM 0.60 20 uM oIMR6436 0.60 1.00 uM 0.60 20 ul oIMR8744 0.20 0.33 ul 0.20 20 uM oIMR8745 0.20 0.33 uM 0.20 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.08 0.03 U/ul 0.08 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 62 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 006618 C57BL/6-Tg(tetO-COX8A/EYFP)1Ksn/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.