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This assay cannot distinguish hemi from hom
Tg= 70 bp
IPC = 74 bp
Tg Sequence:
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
35841 | TTG TGG AAT CAG TGG TTC ATC T | Transgene Forward | A | |||
35842 | CAC TGG AAG ACA AAA CAA ATG C | Transgene Reverse | A | |||
35843 | Fluorophore-1 | AAG GGC TGT GCT GTG GAA TTA | Quencher-1 | Tg Probe | ||
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 | |||
TmoIMR0105 | Fluorophore-2 | CCA ATG GTC GGG CAC TGC TCA A | Quencher-2 | IC Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
35841 | 0.40 uM |
35842 | 0.40 uM |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
Wt Probe | 0.15 uM |
Mutant Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | |
4 | -- | repeat steps 2-3 for 40 cycles | |
5 | 40.0 | -- | Forever |
Stock Number | Strain Name |
---|---|
012253 | B6.Cg-Fxntm1Mkn Tg(FXN)YG8Pook/J |
008586 | B6.Cg-Tg(FXN*)1Sars Fxntm1Mkn/J |
031007 | STOCK Fxnem2.1Lutzy Tg(FXN)Y47Pook/J |
030395 | STOCK Fxnem2.1Lutzy Tg(FXN)YG8Pook/800J |
030930 | STOCK Fxnem2.1Lutzy Tg(FXN)YG8Pook/J |
024097 | STOCK Fxntm1Mkn Tg(FXN)Y47Pook/J |
030324 | STOCK Tg(FXN)YG8Pook/J |
7 strains use this protocol |