Stock Number: 008695 Strain Name: FVB-Tg(tetO-MET)23Rwng/J

Allele: Tg(tetO-MET)23Rwng Protocol Name: Tg(tetO-MET)23Rwng Method: Standard PCR Version: 1.0 Created: 04-September-2009 Updated: 24-September-2009

Expected Results: internal Control= 200bp Tg= 550bp

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8744 - CAA ATG TTG CTT GTC TGG TG - Internal Positive Control Forward oIMR8745 - GTC AGT CGA GTG CAC AGT TT - Internal Positive Control Reverse oIMR9095 - GTC GAG TAG GCG TGT ACG - - oIMR9099 - GAA TGA CAT TCT GGA TGG GTG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) DNA 2.00 - 2.00 ddH2O 4.38 - 4.38 2.5 mM dNTP 0.96 0.20 mM 0.96 10 X High Fidelity Buffer 1.20 1.00 X 1.20 25 mM MgSO4 0.96 2.00 mM 0.96 20 uM oIMR9095 0.60 1.00 uM 0.60 20 uM oIMR9099 0.60 1.00 uM 0.60 20 oIMR8744 0.20 0.33 0.20 20 uM oIMR8745 0.20 0.33 uM 0.20 DMSO 0.80 - 0.80 5 U/ul High Fidelity Platinum Taq 0.10 0.04 U/ul 0.10

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 se - 3 56 45 s - 4 72 1 min - 5 - - Go to step 2-4, 35 times 6 72 2 min - 7 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 008695 FVB-Tg(tetO-MET)23Rwng/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.