Stock Number: 006303 Strain Name: NOD.FVB-Tg(TcraBDC12-4.1)10Jos/GseJ

Allele: Tg(TcraBDC12-4.1)10Jos Protocol Name: Tg(TcraBDC12-4.1)10Jos Method: Standard PCR Version: 1.1 Created: 25-June -2008 Updated: 11-December -2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 525 bp

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

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6498 - CAA TGA AAA CAT ATG CTC CTA C - - oIMR6499 - CTG CCT CCA CTG TTC GCG CC - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.96 - 3.96 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 oIMR6498 0.60 1.00 uM 0.60 20 uM oIMR6499 0.60 1.00 uM 0.60 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 54 30 sec - 4 72 30 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 006303 NOD.FVB-Tg(TcraBDC12-4.1)10Jos/GseJ 009377 NOD.Cg-Rag1tm1Mom Tg(TcraBDC12-4.1)10Jos Tg(TcrbBDC12-4.1)82Gse/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.