Stock Number: 004779 Strain Name: STOCK Mapttm1(EGFP)Klt/J

Allele: Mapttm1(EGFP)Klt Protocol Name: Mapttm1(EGFP)Klt Method: Melt Curve Analysis Version: 4.0 Created: 14-July -2010 Updated: 14-July -2010

Notes: Melting curve analysis is done using a Roche Light Cycler 480. Expected Results: Mutant Tm = 89.5°C +/- 1.0°C Wild type Tm = 83°C+/- 1.0°C

Gel Image

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Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR0872 - AAG TTC ATC TGC ACC ACC G - Mutant oIMR3092 - CTC AGC ATC CCA CCT GTA AC - Wild type oIMR3093 - CCA GTT GTG TAT GTC CAC CC - Wild type oIMR3999 - TGC TCA GGT AGT GGT TGT CG - Mutant

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 1.60 - 1.60 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 20 uM oIMR3092 0.60 1.00 uM 0.60 20 uM oIMR3093 0.60 1.00 uM 0.60 20 uM oIMR0872 0.60 1.00 uM 0.60 20 uM oIMR3999 0.60 1.00 uM 0.60 DNA 2.00 - 2.00

Cycling  Step # Temp °C Time Note 1 94 5 min - 2 94 30 sec - 3 62 1 min - 4 72 30 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 004779 STOCK Mapttm1(EGFP)Klt/J 004807 B6;129-Psen1tm1Mpm Tg(APPSwe,tauP301L)1Lfa/Mmjax 004808 STOCK Mapttm1(EGFP)Klt Tg(MAPT)8cPdav/J 005491 B6.Cg-Mapttm1(EGFP)Klt Tg(MAPT)8cPdav/J 005864 B6.Cg-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.