Stock Number: 014544 Strain Name: STOCK Tg(tetO-ABL1*P242E*P249E)CPdav/J

Allele: Generic tTA Protocol Name: Generic tTA Method: Melt Curve Analysis Version: 1.0 Created: 01-March -2010 Updated: 01-March -2010

Expected Results: Transgene Tm = 92°C +/- 1.0°C Internal positive control Tm = 82°C+/- 1.0°C

Gel Image

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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 oIMR8746 - CGC TGT GGG GCA TTT TAC TTT AG - Transgene Forward oIMR8747 - CAT GTC CAG ATC GAA ATC GTC - Transgene Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.60 - 2.60 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 ul oIMR8744 0.10 0.17 ul 0.10 20 uM oIMR8745 0.10 0.17 uM 0.10 20 uM oIMR8746 0.60 1.00 uM 0.60 20 uM oIMR8747 0.60 1.00 uM 0.60 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 59 1 min - 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 004758 B6;129S7-Wnt5atm1Amc/J 013156 FVB-Tg(tetO-CDK5R1*)1Vln/J 014153 FVB-Tg(Myh6/tetO-Itpr2)3.11Jmol/J 014544 STOCK Tg(tetO-ABL1*P242E*P249E)CPdav/J 016176 B6(Cg)-Tg(tetO-Per2)2Jt/J 905869 B6.Cg-Tg(tTALap)5Bjd/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.