Stock Number: 007880 Strain Name: B6SJL-Tg(Thy1-Stx1a/EYFP)1Sud/J

Allele: Tg(Thy1-Stx1a/EYFP)1Sud Protocol Name: Tg(Thy1-Stx1a/EYFP)1Sud Method: Standard PCR Version: 2.1 Created: 06-May -2009 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: TG = ~550 bp Heterozygote = 200 bp and ~550 bp Wild type = 200 bp

Gel Image

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR7609 - CCT CCT TGG TCT TCT CAT CG - Reverse oIMR7709 - TGG TGA ACC GCA TCG AGC TG - Forward 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 3.56 - 3.56 10 X AB PCR BufferII 1.20 1.00 X 1.20 25 mM MgCl2 0.96 2.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR7709 0.70 1.17 uM 0.70 20 uM oIMR7609 0.70 1.17 uM 0.70 20 ul oIMR8744 0.10 0.17 ul 0.10 20 uM oIMR8745 0.10 0.17 uM 0.10 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 3 min - 2 94 30 sec - 3 60 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 007880 B6SJL-Tg(Thy1-Stx1a/EYFP)1Sud/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.