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Team:Cambridge/Protocols/Colony PCR
From 2011.igem.org
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Contents |
Colony PCR
protocol description
Theory
How it works
Practice
Composition of 50 μl reaction:
| water | 35.7 μl |
| 10 mM dNTPs | 1 μl |
| 10× NH4 Buffer | 5 μl |
| 50mM MgCl2 | 1 μl |
| forward primer | 2.5 μl |
| reverse primer | 2.5 μl |
| liquid culture | 1.3 μl |
| Taq polymerase | 1 μl |
Settings of PCR machine
| Step 1 | 95°C | 6 min |
| Step 2 | 98°C | 10 s |
| 55°C | 30 s | |
| 72°C | 3 min | |
| Step 3 | 72°C | 5 min |
How to do it in the lab
{Standard layout for procedures is to use:
- <Procedure title - aka what you are doing>
- <step 1>
- <step 2>
- <additional notes/important information regarding the previous step>
the text within the < > is what should be written, don't include < > in actual writeup :P
if in doubt see the gel electrophoresis protocol
}
Safety
The safety implication of the procedure.
