Stock Number: 003135 Strain Name: C57BL/6-Tg(TRAMP)8247Ng/J

Allele: Tg(TRAMP)8247Ng Protocol Name: Tg(Tramp1) Method: Standard PCR Version: 1.1 Created: 15-April -2009 Updated: 30-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene =~1200 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 oIMR7082 - CCG GTC GAC CGG AAG CTT CCA CAA GTG CAT TTA - Transgene oIMR7083 - AGG CAT TCC ACC ACT GCT CCC ATT CAT C - Transgene

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.87 - 3.87 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 oIMR7082 0.60 1.00 uM 0.60 20 uM oIMR7083 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 0.15 0.06 U/ul 0.15 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 003135 C57BL/6-Tg(TRAMP)8247Ng/J 008215 (C57BL/6-Tg(TRAMP)8247Ng/J X FVB/NJ)F1/J 308215 C57BL/6-Tg(TRAMP)8247Ng/J X FVB/NJ)F1 Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.