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>chr8:86488607+86488735 129bp CCCTCACAAGCTGATGTCAC TCTCCTGTGGGCCTAACTTG
Mutant= 148 bp
Wild Type = 129 bp
Wt Sequence: ccctcacaagctgatgtcacaggactcaagtcagccTCtactatatggttcctctcggctactgagaggcctaattttcccgggagggccagagtcaaggtcacttggccaagttaggcccacaggaga
Mutant Sequence: ccctcacaagctgatgtcacaggactcaagtcagccTCtgaaagcacactggttttggcagtcaagtttaagttcaaatccttaaacttggactggtgacaaaactactgtaatcccagctacttaggatgtagaagcaggaagacca
526 bp deletion spanning ENSMUSE00001257874 (exon 2) beginning at Chromosome 8 positive strand position 83,964,745 bp CTACTATATGGTTCCTCTCG, and ending after CTAGAACCTTCCTCTTTTAC at 83,965,270 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
35236 | Fluorophore-1 | TCG GCT ACT GAG AGG CCT AAT | Quencher-1 | WT Probe | ||
35237 | CCC TCA CAA GCT GAT GTC AC | Common | A | |||
35238 | TCT CCT GTG GGC CTA ACT TG | Wild type Reverse | A | |||
35239 | TGG TCT TCC TGC TTC TAC ATC C | Mutant Reverse | A | |||
35240 | Fluorophore-2 | ACA CTG GTT TTG GCA GTC AAG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
35237 | 0.40 uM |
35238 | 0.40 uM |
35239 | 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 |