Stock Number: 002226 Strain Name: C57BL/6J-Tg(Alb1HBV)44Bri/J

Allele: Tg(Alb1HBV)44Bri Protocol Name: Tg(Alb1HBV)44Bri Method: Standard PCR Version: 1.1 Created: 25-April -2003 Updated: 24-September-2009

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 223 bp Internal positive control = 113 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 oIMR0064 - AAC ATG GAG AAC ATC ACA TC - Transgene oIMR0065 - AGC GAT AAC CAG GAC AAG TT - Transgene oIMR0066 - ATG TAC TGG TCC CGC ATG GC - - oIMR0067 - TTT GCA GGA CTC CTA CCG G - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.52 - 2.52 10 X PCR Buffer II (ABI) 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 oIMR0064 0.60 1.00 uM 0.60 20 uM oIMR0065 0.60 1.00 uM 0.60 20 uM oIMR0066 0.60 1.00 uM 0.60 20 uM oIMR0067 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 55 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 002226 C57BL/6J-Tg(Alb1HBV)44Bri/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.