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Mutant = 95 bp
Wild type = 93 bp
>chr9:112912424+112912516 93bp TGCAGATATTCACAACCAATCA GGTTACAATCCCCTTCAGCTC
Wt Sequence: AGCAGAGCCTGAAGGAAAGGTCATCCAGAGACTGTCCCACTGTGTGATCCATTCCATCAGCAGACACCAAACCCCGACACTATTACTGATGCTTAGATGTGCTTGAAGACAGAAGCCTGGTATGGCTGTCCTCTGAGAGGGTCTGCCATCACCTGACTAAGACATATGCAGATATTCACAACCAATCATTGgCCTGAGCCTGAGGACCTTAATGGAAGAGTTAGGGAAAGGGATgaAAGAGCTGAAGGGGATTGTAACCCCATAGAAAGAACAAGAGTATCAACTAACTGGACCCCTCAGAGATCCCAGAGACTAAACCACCAACCAAAGAG
Mut Sequence: GCAAAATGGAGCAGTATTTGAACATGGTAGAGTAAGCGAGAACACGTTTGATGTCGGTCTGTACCAGCGCGGCAAAACCGGCCAGCAGCAGCGTaaagagctgaaggggattgtaaccccatagaaagaacaagagtatcaactaactggacccctcagagatcccagagac
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
42433 | ATG GTA GAG TAA GCG AGA ACA CG | Mutant Forward | A | vector | ||
43137 | GGT TAC AAT CCC CTT CAG CTC | Common | A | mChr9 | ||
43138 | TGC AGA TAT TCA CAA CCA ATC A | Wild type Forward | A | mChr9 | ||
43139 | Fluorophore-1 | ATG TCG GTC TGT ACC AGC GCG | Quencher-1 | MUT Probe | ||
43140 | Fluorophore-2 | CCT GAG CCT GAG GAC CTT AAT GG | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
42433 | 0.40 uM |
43137 | 0.40 uM |
43138 | 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 |