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Mutant = 286 and 273 bp
Heterozygote = 286, 273 and 541 bp
Wild type = 541 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
10504 | ACC TAT CAG GCA AAA GGA CG | Forward | ||||
10505 | AGG GAT GGT GAT TGG TTG GA | Reverse |
Component | Final Concentration |
---|---|
ddH2O | |
KAPA Rxn Buffer Contains 1.5 mM Mg2+ final concentration. | 1.00 |
dNTP | 0.20 |
10504 | 1.00 |
10505 | 1.00 |
DNA Loading Dye | 0.69 |
KAPATaq DNA Polymera No proofreading function. 1 error per 40000 nucleotides. Initial denature 2 min. | 0.03 |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | |
4 | 72.0 | -- | |
5 | -- | Go to step 2-4, 35 times | |
6 | 72.0 | -- | |
7 | 10.0 | -- | hold |
Component | Final Concentration |
---|---|
ddH2O | |
NEB buffer 4 | 1.50 |
BspHI | 1.25 |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 37.0 | -- | |
2 | 4.0 | -- | hold |