Stock Number: 002934 Strain Name: FVB.Cg-Tg(Wnt1)1Hev/J

Allele: Tg(Wnt1)1Hev Protocol Name: Tg(Wnt1)1Hev Method: Standard PCR Version: 1.1 Created: 29-August -2000 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 440 bp Internal positive control = 300 bp

Gel Image

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Gel Information: Separate digested products on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0513 - GGA CTT GCT TCT CTT CTC ATA GCC - Transgene oIMR0514 - CCA CAC AGG CAT AGA GTG TCT GC - Transgene oIMR0515 - GCT ATC CAG AAG AAA CCC CTC AAA TCC - - oIMR0516 - CAT GTC TCG ATC CCA GTA GAC GGT C - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.26 - 3.26 10 X PCR Buffer II (ABI) 1.20 1.00 X 1.20 25 mM MgCl2 1.68 3.50 mM 1.68 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR0513 0.30 0.50 uM 0.30 20 uM oIMR0514 0.30 0.50 uM 0.30 20 uM oIMR0515 0.30 0.50 uM 0.30 20 uM oIMR0516 0.30 0.50 uM 0.30 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.04 0.02 U/ul 0.04 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 1.5 min - 2 94 30 sec - 3 58 30 sec - 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 002870 B6SJL-Tg(Wnt1)1Hev/J 002934 FVB.Cg-Tg(Wnt1)1Hev/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.