Genotyping Protocols Database
Protocol 34650: Probe Assay - Generic APP human genomic or cDNA
Version 6.0

Notes

The genotyping protocol(s) presented here have been optimized for reagents and conditions used by The Jackson Laboratory (JAX). To genotype animals, JAX recommends researchers validate the assay independently upon receipt of animals into their facility. Reaction cycling temperature and times may require additional optimization based on the specific genotyping reagents used.

Expected Results

JAX Protocol

Protocol Primers

Primer 5' Label Sequence 5' → 3' 3' Label Primer Type Reaction Note
oIMR1544 CAC GTG GGC TCC AGC ATT Internal Positive Control Forward A
oIMR3580 TCA CCA GTC ATT TCT GCC TTT G Internal Positive Control Reverse A
oIMR5260 TGG GTT CAA ACA AAG GTG CAA Transgene Forward A
oIMR7707 GAT GAC GAT CAC TGT CGC TAT GAC Transgene Reverse A
TmoIMR0076 Fluorophore-1 CAT TGG ACT CAT GGT GGG CGG TG Quencher-1 Tg Probe
TmoIMR0107 Fluorophore-2 CCA ATG GTC GGG CAC TGC TCA A Quencher-2 IC Probe

Reaction A

Component Final Concentration
Kapa Probe Fast QPCR 1.00 X
ddH2O
oIMR1544 0.40 uM
oIMR3580 0.40 uM
oIMR5260 0.40 uM
oIMR7707 0.40 uM
Wt Probe 0.15 uM
Mutant Probe 0.15 uM
DNA

Cycling

Step Temp °C Time Note
1 95.0 --
2 95.0 --
3 60.0 -- repeat steps 2-3 for 40 cycles
JAX uses a very high speed Taq (~1000 bp/sec), use cycling times recommended for your reagents.

Strains Using This Protocol

Stock Number Strain Name
006409 129S1.129(Cg)-Tg(APPSw)40Btla/Mmjax
031988 129S4.Cg-Tg(APPSwe,tauP301L)1Lfa Psen1tm1Mpm/LfaJ
006555 A.129(B6)-Tg(APPSw)40Btla/Mmjax
005300 B6.129-Tg(APPSw)40Btla/Mmjax
005301 B6.129S2-Tg(APP)8.9Btla/J
006406 B6.129S4-Tg(APPSwLon)96Btla/Mmjax
004661 B6.Cg-Zbtb20Tg(PDGFB-APPSwInd)20Lms/1J
008730 B6.Cg-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax
004662 B6.Cg-Tg(PDGFB-APP)5Lms/J
006554 B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax
007027 C57BL/6-Tg(Thy1-APPSwDutIowa)BWevn/Mmjax
006472 D2.129(B6)-Tg(APPSw)40Btla/Mmjax
12 strains use this protocol