Table 1.
Mutation | Cell line | Primers (introduced mutations are underlined) | Primer concentrations | Annealing temperature |
---|---|---|---|---|
c.34 G > A |
A549 |
Mutation specific forward: |
400 nM |
57°C |
|
|
GAATATAAACTTATGGTAGTTGGAGATA |
|
|
|
|
Overlapping forward: |
100 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.34 G > T |
NCI-H23 |
Mutation specific forward: |
400 nM |
60°C |
|
|
GAATATAAACTTGTAGTAATTGGAGCTT |
|
|
|
|
Overlapping forward: |
150 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.34 G > C |
PSN1 |
Mutation specific forward: |
400 nM |
60°C |
|
|
GAATATAAACTTGTAGTAATTGGAGCTC |
|
|
|
|
Overlapping forward: |
100 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.35 G > A |
LS174T |
Mutation specific forward: |
400 nM |
60°C |
|
|
GAATATAAACTTGTGGTAATTGGAGATGA |
|
|
|
|
Overlapping forward: |
150 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.35 G > T |
SW480 |
Mutation specific forward: |
400 nM |
64°C |
|
|
GAATATAAACTTGTAGTAATTGGAGCTGT |
|
|
|
|
Overlapping forward: |
150 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.35 G > C |
RPMI8226 |
Mutation specific forward: |
400 nM |
58°C |
|
|
GAATATAAACTTGTAGTAATTGGAGCTGC |
|
|
|
|
Overlapping forward: |
100 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
|
|
ACTGTCAAGGCACTCTTGCCTAC |
|
|
c.38 G > A |
DLD-1 |
Mutation specific forward: |
400 nM |
62°C |
|
|
AAACTTGTGGTAGTTGGAGATGGTTA |
|
|
|
|
Overlapping forward: |
150 nM |
|
|
|
ATGACTGAATATAAACTTGTGGTAGTTG |
|
|
|
|
Common reverse: |
400 nM |
|
ACTGTCAAGGCACTCTTGCCTAC |
Primer sequences are given in 5’ → 3’ directions.