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H2-T18<b> Variant = (Jr 457) = "hom"
H2-T18<b>= (Jrs 666, 664, 438) = "wt"
Please run JR 683 as well at validation
664 and 457 controls must be run 1-18-16 ESP
(T/C)TG(GT/AC)GTTCTGATGAGTTATTGATCTAG(C/T)TATGATAATCAGTGATTG(ATTG/----)CTATT(A/G)T(A/C)CACAGTCACAAGCTCAAGATTATGAAT(T/C)TCCTA(C/T)G(C/G)GTTGTCTCTAAGTATA(TA/--)GACTTAAAAGGGCTTCTCATTGAGCT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
25550 | Fluorophore-1 | AAT CAG TGA TTG ATT GCT ATT ATA AC | Quencher-1 | WT Probe | A | |
26436 | Fluorophore-2 | TCA GTG ATT GCT ATT GTC ACA GTC | Quencher-2 | MUT Probe | A | |
26747 | TGA CGT TCT GAT GAG TTA TTG ATC T | Forward | A | |||
26748 | AGC TCA ATG AGA AGC CCT TT | Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
25550 | 0.40 uM |
26436 | 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 |