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This mutation introduces a DdeI enzyme restriction site.
Mutant = C/C
WT = G/G
Het = C/G
ATGTTCTTGGCTGTTTTGTATTGCCTTCTGTGGAGTTTCCAGATCTCTGATGGCCATTTTCCTCGAGCCTGTGCCTCCTCTAAGAACTTGTTGGCAAAAGAATG
CTGCCCACCATGGATGGGTGATGGGAGTCCCTGCGGCCAGCTTTCAGGCAGAGGTTCCTGCCAGGATATCCTTCTGTCCAGTGCACCATCTGGACCTCAGTTCCCCTTCAAAGGGGTGGA
TGACCGTGAGTCCTGGCCCTCTGTGTTTTATAATAGGACCTGCCAGTGCTCAGGCAACTTCATGGGTTTCAACTGCGGAAACT(g/c)TAAGTTTGGATTT
GGGGGCCCAAATTGTACAGAGAAGCGAGTCTTGATTAGAAGAAACATTTTTGATTTGAGTGTCTCCGAAAAGAATAAGTTCTTTTCTTACCTCACTTTAGCAAAACATACTATCAGCTCAGTCT
ATGTCATCCCCACAGGCACCTATGGCCAAATGAACAATGGGTCAACACCCATGTTTAATGATATCAACATCTACGACCTCTTTGTATGGATGCATTACTATGTGTC
Albino = C
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17285 | AAT AGG ACC TGC CAG TGC TC | Forward | A | |||
17286 | GCC TGT GGG GAT GAC ATA GA | Reverse | A | |||
17430 | Fluorophore-1 | CTG CGG AAA CTC TAA GTT TGG AT | Quencher-1 | MUT Probe | A | |
17432 | Fluorophore-2 | CTG CGG AAA CTG TAA GTT TGG AT | Quencher-2 | WT Probe | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
17285 | 0.40 uM |
17286 | 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 |