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Team:Heidelberg/Templates/Methods
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Colonies were picked with pipet tips and dipped into the PCR mixture. | Colonies were picked with pipet tips and dipped into the PCR mixture. | ||
Revision as of 14:15, 4 October 2013
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 |
