Stock Number: 006472 Strain Name: D2.129(B6)-Tg(APPSw)40Btla/J

Allele: Tg(APPSw)40Btla Protocol Name: Tg(APPSw)40Btla Method: Melt Curve Analysis Version: 3.0 Created: 27-May -2009 Updated: 24-September-2009

Notes: Melting curve analysis is done using a Roche Light Cycler 480. This assay will NOT distinguish hemizygous from homozygous transgenic animals. Expected Results: Tg/O = lower curve WT = Higher curve

Gel Image

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Gel Information: .

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR6938 - CTT CAC TCG TTC TCA TTC TCT TCC A - Transgene oIMR6939 - GCG TTT TTA TCC GCA TTT CGT TTT T - 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 0.64 - 0.64 10 X High Resolution Melt Master Mix (Roche) 6.00 5.00 X 6.00 20 uM oIMR6938 0.60 1.00 uM 0.60 20 uM oIMR6939 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 DNA 2.00 - 2.00 25 mM MgCl2 0.96 2.00 mM 0.96

Cycling  Step # Temp °C Time Note 1 94 10 min - 2 94 30 sec - 3 60 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 006472 D2.129(B6)-Tg(APPSw)40Btla/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.