Stock Number: 002709 Strain Name: B6;C3-Tg(TettTALuc)1Dgs/J

Allele: Tg(TettTALuc)1Dgs Protocol Name: Tg(luc) Method: Standard PCR Version: 1.1 Created: 23-October -2006 Updated: 24-September-2009

Expected Results: Transgene = 458 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 oIMR4979 - GCG GAA TAC TTC GAA ATG TCC - Transgene oIMR4980 - CCT TAG GTA ACC CAG TAG ATC C - Transgene oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 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 oIMR4979 0.60 1.00 uM 0.60 20 uM oIMR4980 0.60 1.00 uM 0.60 20 oIMR8744 0.30 0.50 0.30 20 uM oIMR8745 0.30 0.50 uM 0.30 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 58 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 002709 B6;C3-Tg(TettTALuc)1Dgs/J 006098 B6.Cg-Tg(Il2/NFAT-luc)83Rinc/J 006100 B10.Cg-H2k Tg(NFkB/Fos-luc)26Rinc/J 006101 B6.Cg-Tg(TRE/Prl-luc)31FlvRinc/J 006102 B10.Cg-H2k Tg(Il2/NFAT-luc)83Rinc/J 007800 FVB/N-Tg(Ins1-luc)VUPwrs/J 904292 MBP.luc Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.