Team:Heidelberg/Templates/Methods
From 2013.igem.org
Colony PCR
Colonies were picked with pipet tips and dipped into the PCR mixture.
Ingredient | amount [µl] |
---|---|
Overnight Culture | Top of a pipet tip |
10x PCR buffer | 2.0 |
MgCl2 (25mM) | 2.0 |
Forward primer (10µM) | 0.2 |
Reverse primer (10µM) | 0.2 |
dNTPs (10mM) | 1.0 |
Polymerase | 0.5 |
Sterile water | ad 10.0 |
Total | 10.0 |
The following PCR program was adjusted according to annealing conditions.
Cycles | temperature [°C] | Time [min:s] |
---|---|---|
1 | 98 | 2:00 |
35 | 98 | 0:05 |
68 (annealing) | 0:15 | |
72 | 1:00 per kbp | |
1 | 72 | 10:00 |
1 | 12 | inf |