Stock Number: 006770 Strain Name: STOCK Rag1tm1Mom Tg(TIE2GFP)287Sato/J

Allele: Tg(TIE2GFP)287Sato Protocol Name: Tg(TIE2GFP)287Sato Method: Standard PCR Version: 1.1 Created: 08-September-2000 Updated: 30-August -2011

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 567 bp Internal positive control=324 bp

Gel Image

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Gel Information: Separate by gel electrophoresis on a 1.5% agarose gel.

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR1263 - ATT CTC GTG GAA CTG GAT GG - Transgene oIMR1264 - GGA CAG GTA ATG GTT GTC TGG - Transgene oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.36 - 3.36 10 X AB PCR BufferII 1.20 1.00 X 1.20 25 mM MgCl2 1.20 2.50 mM 1.20 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR1263 0.60 1.00 uM 0.60 20 uM oIMR1264 0.60 1.00 uM 0.60 20 uM oIMR7338 0.30 0.50 uM 0.30 20 uM oIMR7339 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 0.06 0.03 U/ul 0.06 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 1.5 min - 2 94 30 sec - 3 58 1 min - 4 72 1 min 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 003658 STOCK Tg(TIE2GFP)287Sato/J 004659 B6.Cg-Tg(TIE2GFP)287Sato/1J 006207 STOCK Tg(Pcp2-cre)1Amc/J 006770 STOCK Rag1tm1Mom Tg(TIE2GFP)287Sato/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.