Stock Number: 003528 Strain Name: STOCK Tg(GFAP-TVA)5Hev/J

Allele: Tg(GFAP-TVA)5Hev Protocol Name: Tg(GFAP-TVA)5Hev, Tg(NES-TVA)12Hev Method: Standard PCR Version: 1.1 Created: 28-April -2005 Updated: 04-January -2012

Expected Results: TG = ~500 bp internal control = ~398 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 oIMR1111 - CTG CTG CCC GGT AAC GTG ACC GG - - oIMR1112 - GCC CTG GGG AAG GTC CTG CCC - - oIMR3172 - AGA GCT CCG GGT TCT CTC TC - Common oIMR3173 - GGG AGG AAG GAA CTC CAC TC - Wild type

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.77 - 2.77 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 oIMR1111 0.60 1.00 uM 0.60 20 uM oIMR1112 0.60 1.00 uM 0.60 20 uM oIMR3172 0.60 1.00 uM 0.60 20 uM oIMR3173 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 0.05 0.02 U/ul 0.05 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 66 1 min - 4 72 1 min repeat steps 2-4 for 35 cycles 5 72 10 min - 6 10 - hold

This genotyping protocol is used for the following strains:   Stock Number Strain Name 003528 STOCK Tg(GFAP-TVA)5Hev/J 003529 STOCK Tg(NES-TVA)J12Ech/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.