Stock Number: 004662 Strain Name: B6.Cg-Tg(PDGFB-APP)5Lms/J

Allele: Tg(PDGFB-APP)5Lms Protocol Name: Tg(PDGFB-APP) Method: Standard PCR Version: 1.1 Created: 10-October -2003 Updated: 24-September-2009

Expected Results: Transgene = 360 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 oIMR2044 - GGT GAG TTT GTA AGT GAT GCC - Transgene oIMR2045 - TCT TCT TCT TCC ACC TCA GC - Transgene 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 2.79 - 2.79 10 X PCR Buffer II (ABI) 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 oIMR2044 0.60 1.00 uM 0.60 20 uM oIMR2045 0.60 1.00 uM 0.60 20 oIMR8744 0.60 1.00 0.60 20 uM oIMR8745 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 0.03 0.01 U/ul 0.03 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 1.5 min - 2 94 30 sec - 3 62 1 min - 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 004661 B6.Cg-Tg(PDGFB-APPSwInd)20Lms/1J 004662 B6.Cg-Tg(PDGFB-APP)5Lms/J 006293 B6.Cg-Tg(PDGFB-APPSwInd)20Lms/2J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.