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Transgene = 377 bp
Internal positive control = 324 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR3610 | AGG ACT GAC CAC TCG ACC AG | Transgene Forward | A | APP | ||
oIMR3611 | CGG GGG TCT AGT TCT GCA T | Transgene Reverse | A | APP | ||
oIMR7338 | CTA GGC CAC AGA ATT GAA AGA TCT | Internal Positive Control Forward | A | |||
oIMR7339 | GTA GGT GGA AAT TCT AGC ATC ATC C | Internal Positive Control Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
oIMR3610 | 0.50 uM |
oIMR3611 | 0.50 uM |
oIMR7338 | 0.50 uM |
oIMR7339 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |
Stock Number | Strain Name |
---|---|
005866 | B6.Cg-Tg(APP695)3Dbo Tg(PSEN1dE9)S9Dbo/Mmjax |
006006 | B6.Cg-Tg(Prnp-APP)A-2Dbo/J |
006005 | B6.Cg-Tg(Prnp-App/APPswe)E1-2Dbo/Mmjax |
007051 | B6.Cg-Tg(tetO-APPSwInd)102Dbo/Mmjax |
007049 | B6.Cg-Tg(tetO-APPSwInd)885Dbo/Mmjax |
004807 | B6;129-Tg(APPSwe,tauP301L)1Lfa Psen1tm1Mpm/Mmjax |
004462 | B6;C3-Tg(APPswe,PSEN1dE9)85Dbo/Mmjax |
006004 | B6C3-Tg(tetO-APPSwInd)885Dbo/Mmjax |
003375 | C3B6-Tg(APP695)3Dbo/Mmjax |
007027 | C57BL/6-Tg(Thy1-APPSwDutIowa)BWevn/Mmjax |
035561 | STOCK Tc(HSA21,CAG-EGFP)1Yakaz/J |
11 strains use this protocol |