Stock Number:008730
Strain Name:B6.Cg-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax
Generic Human PSEN1 cDNA
Generic Human PSEN1 cDNA Multiplex1
Melt Curve Analysis
1.0
07-February -2012 09-February -2012
Notes:

This assay will NOT distinguish hemizygous from homozygous transgenic animals.
Expected Results:
Internal Control- TM- 82 delta 2 Area- 10
Transgene- TM-  79 delta 2 Area- 10
Gel Image 
Image Image file
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Protocol Primers 
Primer 5' Label Sequence 5' --> 3' 3' Label Primer Type
13763 - TCA TGA CTA TCC TCC TGG TGG - Transgene Forward
13764 - CGT TAT AGG TTT TAA ACA CTT CCC C - Transgene Reverse
oIMR7338 - CTA GGC CAC AGA ATT GAA AGA TCT - Internal Positive Control Forward
oIMR7339 - GTA GGT GGA AAT TCT AGC ATC ATC C - Internal Positive Control Reverse
Reaction/Components 
Reaction Component Volume (µl) Final
Concentration		 Total Volume (µl)
ddH2O 5.45 - 5.45
5 X Kapa 2G HS buffer 2.40 1.00 X 2.40
25 mM MgCl2 0.96 2.00 mM 0.96
10 mM dNTPS-kapa 0.24 0.20 mM 0.24
10 uM 13764 0.30 0.25 uM 0.30
10 uM 13763 0.30 0.25 uM 0.30
2.5 ul Kapa 2G HS taq polymerase 0.05 0.01 ul 0.05
DNA 2.00 - 2.00
20 uM oIMR7339 0.15 0.25 uM 0.15
20 uM oIMR7338 0.15 0.25 uM 0.15
Cycling 
Step # Temp °C Time Note
1 94 2 min -
2 94 20sec -
3 65 15sec -1.5 C per cycle
4 68 10sec -
5 - - repeat step 2-4 for 10 cycles
6 94 15sec -
7 50 15sec -
8 72 10sec -
9 - - repeat step 6-8 for 28 cycles
10 72 2 min -
11 10 - hold
This genotyping protocol is used for the following strains:
 
Stock Number Strain Name
005491 B6.Cg-Mapttm1(EGFP)Klt Tg(MAPT)8cPdav/J
005864 B6.Cg-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax
005866 B6.Cg-Tg(APP695)3Dbo Tg(PSEN1dE9)S9Dbo/Mmjax
006554 B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax
008730 B6.Cg-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax
Genotyping resources and troubleshooting
This page contains guidelines, frequently asked questions, and helpful external links to genotyping and PCR resources.