For in-depth product & services help, ask our
Technical Information Scientists
Ref Seq:AGGAGATCGAGGTGGAGGTATTTG(A/C)
Mut = 29 bases highlighted in R chromat
WT = A (reportedly)
~325 bp
GTGATAGTCCACGAGGTTCCAGGACAGGAGATCGAGGTGGAGGTATTTG(A/C)CAA(GGA/ATC)TCCAGATAAA(GA/TC)TG(A/C)TTT(T/C)CT(G/A)GGAAG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
24240 | GAC AGG AGA TCG AGG TGG AG | Forward | A | |||
24241 | TGT GAG TGC AGG TGC CTA AG | Reverse | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
24240 | 0.50 uM |
24241 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |