Magni:Protocols/Inocula
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(New page: {{Magni}} <div style="padding: 10px; width: 720px; border: 5px solid #000000;"> = Plasmid propagation = To propagate a plasmid ''in vivo'', infect selective liquid medium with a single co...) |
Current revision (16:42, 9 September 2012) (view source) (→Plasmid propagation) |
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To propagate a plasmid ''in vivo'', infect selective liquid medium with a single colony from a transformation plate. | To propagate a plasmid ''in vivo'', infect selective liquid medium with a single colony from a transformation plate. | ||
| + | ALWAYS WORK NEAR THE FLAME! | ||
*Prepare 5 ml of LB + antibiotic in a 15-ml tube. | *Prepare 5 ml of LB + antibiotic in a 15-ml tube. | ||
*Select a well-isolated colony from a transformation plate. | *Select a well-isolated colony from a transformation plate. | ||
Current revision
Plasmid propagation
To propagate a plasmid in vivo, infect selective liquid medium with a single colony from a transformation plate.
ALWAYS WORK NEAR THE FLAME!
- Prepare 5 ml of LB + antibiotic in a 15-ml tube.
- Select a well-isolated colony from a transformation plate.
- Take a P20 with a sterile tip.
- Touch this colony with the tip and infect the liquid medium.
- Close the 15-ml tube and incubate it at 37degC, 220 rpm overnight (14-24 hours).
- Seal the transformation plate with parafilm and store in the fridge.
NOTES
- Heat-sensitive plasmids must be incubated at a proper temperature. If an incubator set at a low temperature is not available, incubate the culture at room temperature on the rotating mixer with rolls.
