Stock Number: 006554 Strain Name: B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/J

Allele: Pde6brd1-1H Protocol Name: Generic Pde6b Method: Melt Curve Analysis Version: 3.0 Created: 30-June -2009 Updated: 24-September-2009

Notes: Melting curve analysis is done using a Roche Light Cycler 480. Expected Results: Mutant Tm = 85°C +/- 1.0°C Wild type Tm = 81.5°C+/- 1.0°C

Gel Image

Image Image file View Full Size

Protocol Primers

Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type oIMR2093 - AAG CTA GCT GCA GTA ACG CCA TTT - Mutant oIMR2094 - ACC TGC ATG TGA ACC CAG TAT TCT ATC - Wild type oIMR2095 - CTA CAG CCC CTC TCC AAG GTT TAT AG - Common

Number of Reactions

Reaction/Components  Reaction Component Volume (µl) Final Concentration Total Volume (µl) ddH2O 2.20 - 2.20 2 X SYBR Green Master Mix (Roche) 6.00 1.00 X 6.00 DNA 2.00 - 2.00 20 uM oIMR2093 0.60 1.00 uM 0.60 20 uM oIMR2094 0.60 1.00 uM 0.60 20 uM oIMR2095 0.60 1.00 uM 0.60

Cycling  Step # Temp °C Time Note 1 94 7.5 min - 2 94 30 sec - 3 55 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 001924 B6EiC3Sn a/A-Ts(1716)65Dn 002810 B6CBA-Tg(HDexon1)62Gpb/1J 004462 B6C3-Tg(APPswe,PSEN1dE9)85Dbo/J 004808 STOCK Mapttm1(EGFP)Klt Tg(MAPT)8cPdav/J 004850 B6EiC3Sn-Rb(12.Ts171665Dn)2Cje/CjeDn 004861 B6EiC3Sn-Ts(16C-tel)1Cje 005965 STOCK Tg(Pomc1-cre)16Lowl/J 006494 B6CBA-Tg(HDexon1)62Gpb/3J 006554 B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/J 006576 B6.FVB-Tg(GNAT2-Dta)98Wwk/J Genotyping resources and troubleshooting This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.