Stock Number: 008596 Strain Name: B6.Cg-Tg(Prnp-Abca1)EHol/J

Allele: Tg(Prnp-Abca1)EHol Protocol Name: Tg(Prnp-Abca1) Method: Standard PCR Version: 3.0 Created: 09-September-2011 Updated: 09-September-2011

Notes: This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Transgene = 765 bp Internal positive control = 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 oIMR8744 - CAA ATG TTG CTT GTC TGG TG - - oIMR8745 - GTC AGT CGA GTG CAC AGT TT - - oIMR9291 - CCT GCA GCA CAA TTT GTC CCT TCC AAG ATC - - oIMR9292 - AGG TCA TGG AAC ACA GCC AAC TCC TGA AAG - -

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 3.78 - 3.78 10 X AB PCR BufferII 1.20 1.00 X 1.20 50 mM MgCl2 0.96 4.00 mM 0.96 2.5 mM dNTP 0.96 0.20 mM 0.96 20 uM oIMR9291 0.60 1.00 uM 0.60 20 uM oIMR9292 0.60 1.00 uM 0.60 20 ul oIMR8744 0.08 0.13 ul 0.08 20 uM oIMR8745 0.08 0.13 uM 0.08 5 mM DNA Loading Dye 1.66 0.69 mM 1.66 5 U/ul Taq DNA Polymerase 0.08 0.03 U/ul 0.08 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 3 min - 2 94 30 sec - 3 60 45 sec - 4 72 45 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 008596 B6.Cg-Tg(Prnp-Abca1)EHol/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.