Stock Number: 008226 Strain Name: B6.FVB-Tg(CAG-EGFP,-ALPP)2.6Ggc/J

Allele: Tg(CAG-EGFP,-ALPP)2.6Ggc Protocol Name: Tg(CAG-EGFP,-ALPP) Method: Melt Curve Analysis Version: 1.0 Created: 26-May -2009 Updated: 24-September-2009

Expected Results: Transgene Tm = 90.1°C +/- 1.0°C Internal positive control Tm = 82.2°C+/- 1.0°C

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8694 - AGC CAC AAC GTC TAT ATC ATG - Forward oIMR8695 - AAT TTC GAG ATC TGG ATC CTC - Reverse 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 2.00 - 2.00 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR8694 0.60 1.00 uM 0.60 20 uM oIMR8695 0.60 1.00 uM 0.60 20 ul oIMR8744 0.40 0.67 ul 0.40 20 uM oIMR8745 0.40 0.67 uM 0.40 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 15 sec - 3 58 1 min - 4 72 30 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 008200 FVB/N-Tg(CAG-EGFP,-ALPP)2.6Ggc/J 008226 B6.FVB-Tg(CAG-EGFP,-ALPP)2.6Ggc/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.