Stock Number: 002933 Strain Name: FVB/NJ-Tg(MMTVTGFB1)46Hlm/J

Allele: Tg(MMTVTGFB1)46Hlm Protocol Name: Tg(MMTVTGFB1)46Hlm Method: Standard PCR Version: 2.1 Created: 20-August -2008 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 339 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 oIMR1301 - TGC CTA TCA GAA GGT GGT GG - Transgene Forward oIMR1302 - ATA AGG AAT GGA CAG CAG GG - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.99 - 3.99 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 oIMR1301 0.60 1.00 uM 0.60 20 uM oIMR1302 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 0.03 0.01 U/ul 0.03 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 1.5 min - 2 94 30 sec - 3 55 45 sec - 4 72 45 sec 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 002375 B6;D2-Tg(MMTVTGFB1)46Hlm/J 002933 FVB/NJ-Tg(MMTVTGFB1)46Hlm/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.