Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= 118 bp
Wild Type = 111 bp
>chr7:99470282+99470392 111bp AGCCTGCCTGGTTATACACG GGTGGGTGGGAAACTAGAGG
Wt Sequence (deletion in lower case):
AGTCATATTCAAAACTAAGCGTGACTCTGAGAGCCCGTAATCCATAAGCCTCAGTTTTCTCCTCTGTAGAAGGCGCAATAGTCCTAACAACCATCTGAAGGACTAAACAACAAAAGGGGAAGAAAGGCTTTTTCCCCCTGTAACTTAGAGGCCAGCAGACTGGGGTAAGAGTTAGTGACTTCTTCCTGTCTGTAACCAGAGGTCAGGATGGAAGTGGTGGCTTCCAGGGGCAGCTGCCAGGGTAGGCAGAAAGCCAGCGgcattgtctggtgtgatcaaaacagaaaggacctgagttggaagcctgtaaatccagtgccagtgccgagtaagcagaaacaggatagggagttcgagaacagaggctgtagaaaaaacaaacccatgtgctgaggatatagtttagagaagagcacacagcgctctggttcagtaatccccagcaccgtttggggggcgggggcgcacagcgtgagaagaacagcagtgtaggtgtggatctagaggtcttgaatttatccccaccccaggttcatggacttagctgaagtgatcgtggtcattggcgggtgtgaccgaaaggggctcctgaagctgccttttgcggatgcttaccacccagaaagtcagcgctggaccccactgcctagcctgcctggttatacacgctctgaatttgcctcctgtgcacttcgaaatgacatttacgtttctggtgagggtacaagtaaaataggagcctctagtttcccacccacccccttatattttcctccagtgaggagatgggggctggttagtgactaaaactgtgtggtccagcaaagcctagtcaaaatgggagtgctgatgctggggtttgggccgaaggctttaagccttggtgttagtctttgctctgacaagaggctacctggtattcagtcggccattcctggtccctaaactccatcctcatttcctatatggagaccctgtAATAATCCTAATACCTCAGAGAATGAGGATGGAAATGAAGGCAAAGCTTTAGTAAAGAGAGATGACTGTGCCCCGTCCACTCCACCACTGTGCAGACACTCTGCTTTCCCTTGGCCCAACACACATTCTCAGTGACACTTGGGGACACTCCTGCTTTGCTTCCCTGAGCACAAGATGTGTCACACCACAGGC
This mutation is a 719 bp deletion beginning at Chromosome 7 position 99,469,893 bp and ending after 99,470,611 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
68450 | AGC CTG CCT GGT TAT ACA CG | Wild type Forward | A | |||
68451 | GGT GGG TGG GAA ACT AGA GG | Wild type Reverse | A | |||
68452 | TGT AAC CAG AGG TCA GGA TGG | Mutant Forward | A | |||
68453 | AAG CTT TGC CTT CAT TTC CA | Mutant Reverse | A | |||
68455 | Fluorophore-1 | AGG CAG AAA GCC AGC GAA TAA TC | Quencher-1 | MUT Probe | ||
71038 | Fluorophore-2 | CAC TTC GAA ATG ACA TTT ACG TTT | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
68450 | 0.40 uM |
68451 | 0.40 uM |
68452 | 0.40 uM |
68453 | 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 |
We use cookies to personalize our website and to analyze web traffic to improve the user experience. You may decline these cookies although certain areas of the site may not function without them. Please refer to our privacy policy for more information.