Stock Number: 008222 Strain Name: B6.FVB-Tg(IGFBP2)1Miel/J

Allele: Tg(IGFBP2)1Miel Protocol Name: Tg(IGFBP2)1Miel MCA Method: Melt Curve Analysis Version: 1.0 Created: 28-October -2009 Updated: 28-October -2009

Expected Results: Transgene Tm = 89.7°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 oIMR8102 - CCT GTG CAG CAG CTT CTC GCT GAG - - oIMR8103 - CCC AGG CTG GCA CCA TGC TCA CCT G - - oIMR8744 - CAA ATG TTG CTT GTC TGG TG - - oIMR8745 - GTC AGT CGA GTG CAC AGT TT - -

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 oIMR8102 0.60 1.00 uM 0.60 20 uM oIMR8103 0.60 1.00 uM 0.60 20 oIMR8744 0.40 0.67 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 se - 3 65 1 min - 4 72 30 se 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 008222 B6.FVB-Tg(IGFBP2)1Miel/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.