Stock Number: 002373 Strain Name: B6D2-Tg(MMTVTGFA)29Rjc/J

Allele: Tg(MMTVTGFA)29Rjc Protocol Name: Tg(MMTVTGFA) Method: Standard PCR Version: 1.1 Created: 27-October -2000 Updated: 24-September-2009

Expected Results: Transgene = 192 bp

Gel Information: Separated by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0398 - AGT TCT GCT TCC ATG CAA CC - Transgene oIMR0399 - TGA TGA TAA GGA CAG CCA GG - Transgene

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.99 - 3.99 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 oIMR0398 0.60 1.00 uM 0.60 20 uM oIMR0399 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 3 min - 2 94 30 sec - 3 54 30 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 002373 B6D2-Tg(MMTVTGFA)29Rjc/J 002459 B6D2-Tg(MMTVTGFA)254Rjc/J 002953 FVB.Cg-Tg(MMTVTGFA)254Rjc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.