A | B | C | |
Reagent Name | Initial Conc. | Volume (μL) | |
gotaq green 2x | 2X | 5 | |
oligo 1 | 10uM | 0.6 | |
oligo 2 | 10 uM | 0.6 | |
cell suspension in water | cells suspension | 0.8 | |
ddH2O | N/A | 3 | |
TOTAL Volume: | 10 uL | ||
A | B | C | D | |
Number of Cycles | Step Name | Temperature | Duration | |
1 | Initial Denaturation | 98oC | 10min | |
30 | Denaturation | 98oC | 10 sec | |
Annealing | 50-55oC | 30 sec | ||
1min (depends on the length) | 72oC | 1min per 1kb | ||
1 | Hold | 4-10oC | hold |
A | B | C | |
Reagent Name | Initial Conc. | Volume (μL) | |
gotaq green 2x | 2X | 5 | |
oligo 1 | 10uM | 0.6 | |
oligo 2 | 10 uM | 0.6 | |
cell suspension in 20mM | yeast suspension | 0.8 | |
ddH2O | N/A | 3 | |
TOTAL Volume: | 10 uL | ||
A | B | C | D | |
A | B | C | D | |
Number of Cycles | Step Name | Temperature | Duration | |
1 | Initial Denaturation | 98oC | 2min | |
30-33 | Denaturation | 98oC | 10 sec | |
Annealing | 52-55oC | 1min 30sec | ||
1min (depends on the length) | 72oC | 1min per 1kb | ||
1 | Hold | 4-10oC | hold |