This mutation is a 555 bp deletion beginning at Chromosome 10 positive strand position 28,263,339 bp, TTTTCTATGGCCCTGCCCCA, and ending after TGTTTCCCAACTCCTTCTTG at 28,263,893 bp (GRCm38/mm10).
Mutant= 124 bp
Wild Type = 150 bp
Wt genomic sequence with the deletion in uper case
ctgaatccgactcctgcttgtctaccatctgaccctaagttagcagttatgtcccatatttctatattgtaaatcagggttaatatcaagttgtagagatgaataaactgtttttgtctagtttatgaaatgttagtaagaaatggatctacgtgatgtcttgactgatatTTTTCTATGGCCCTGCCCCAGAAGACATTTTAAAATTTAATGTTTTCATAGAGTAATATCCTATAATACTCTATGAAATTTTATAGTTGTAATATTTTACAGTTGTTTGATACCCCTGGCTTATCAATGTTCTTAGCTCTGTTATTTGTAATGTCATTTCAGGTTCCTATATGGTTGTGGACTCCTCAAATCATGATCCTGGAGAAAAAGCCAGACTTCAGCTGCCTACCATGAAGGAGAATGACACCCACTGCATTGATTTCAGTTACCTGTTATATAGCCAGAAGGGGTTGAACCCTGGCACTTTGAATATCCTAGTTAGGGTGAATAAAGGACCTCTTGCTAATCCAATTTGGAATGTAACTGGATTCACTGGTCGTGATTGGCTTCGGGCTGAACTAGCTGTGAGCACCTTTTGGCCCAATGAATACCAGGTAATCCCCTGAGATTTTTCTGGTTTGTGCTGTCTTGTATTGGGTATCTTTGTATATATAGATATAAATACTGGTCCTGCAGGAGTAGGATCCCTCCTTCTTGTTTCCCAACTCCTTCTTGaggaggagtttgagtgccttgacacagaaccttctaactatagaaaatatttccatgcatagaaaacagtagtttggcccacaatttgttgt
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
32631 | GAA ATG GAT CTA CGT GAT GTC TTG | Common | A | |||
32632 | Fluorophore-1 | CCT GCC CCA GAA GAC ATT T | Quencher-1 | WT Probe | ||
32633 | Fluorophore-2 | AGG AGT TTG AGT GCC TTG ACA | Quencher-2 | MUT Probe | ||
32634 | CAG GGG TAT CAA ACA ACT GTA AAA T | Wild type Reverse | A | |||
32635 | ACA ACA AAT TGT GGG CCA AAC | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
32631 | 0.40 uM |
32634 | 0.40 uM |
32635 | 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 |