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Team:Groningen/Template/MODULE/Notebook/protocols/PCR
From 2014.igem.org
Polymerase Chain Reaction
All PCR reactions were done with DreamTaq and Phusion polymerase from Thermo Scientific.
DreamTaq polymerase
For colony PCR DreamTaq polymerase was used. As template, the colony of interest was picked from a plate using a sterile toothpick and mixed into the reaction mixture. The cycling conditions are listed in table 1 below.
| Step | Temperature | Time | Number of cycles |
|---|---|---|---|
| Initial denaturation | 95 °C | 1 - 3 min | 1 |
| Denaturation | 95 °C | 30 sec | 30 |
| Annealing | Tm - 5 °C | 30 sec | |
| Extension | 72 °C | 1 min/kb | |
| Final extension | 72 °C | 10 min/kb | 1 |
