Stock Number: 008168 Strain Name: STOCK Tg(tetO-DTA)1Gfi/J

Allele: Tg(tetO-DTA)1Gfi Protocol Name: tg(teto-DTA)1Gfi/j Method: Melt Curve Analysis Version: 1.0 Created: 14-August -2009 Updated: 24-September-2009

Expected Results: IC- 82 area- 10 Tg/0- Tg-88 area- 10 Tg/0- IC- 82 area- .5

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR8035 - GGC GTG TAC GGT GGG AGG - - oIMR8036 - GGC ATT ATC CAC TTT TAG TGC - - 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 1.60 0.00 1.60 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 oIMR8744 0.60 1.00 0.60 20 uM oIMR8745 0.60 1.00 uM 0.60 DNA 2.00 - 2.00 20 uM oIMR8035 0.60 1.00 uM 0.60 20 uM oIMR8036 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 67 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 008168 STOCK Tg(tetO-DTA)1Gfi/J 008468 B6.Cg-Tg(tetO-DTA)1Gfi/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.