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05/08/13
From 2013.igem.org
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* Protocols from both Bioline kit and Thermoscientific were used for the miniprep. | * Protocols from both Bioline kit and Thermoscientific were used for the miniprep. | ||
* there were 6 tubes containing the plasmid. | * there were 6 tubes containing the plasmid. | ||
| - | * The plasmid concentration in each of the 6 tubes were measured. | + | * The plasmid concentration in each of the 6 tubes were measured using nanodrop. |
* 200ng of plasmid from each of the 6 different tube was used for digestion. | * 200ng of plasmid from each of the 6 different tube was used for digestion. | ||
Revision as of 14:11, 5 August 2013
Isolation of ligated plasmid
- The ligated plasmid- chloramphenicol backbone and limonene biobrick, were isolated from cells that were incubated overnight in broth by miniprep.
- Protocols from both Bioline kit and Thermoscientific were used for the miniprep.
- there were 6 tubes containing the plasmid.
- The plasmid concentration in each of the 6 tubes were measured using nanodrop.
- 200ng of plasmid from each of the 6 different tube was used for digestion.
Digestion of ligated plasmid
- Master Mix (changes were made from the protocol)
- 2ul of NEB Buffer
- 1ul of EcoRI
- 1ul of PstI
- varying concentration of dH2O and plasmid depending on overall concentration of plasmid in each tube.
- Final volume of each of the 6 tubes was 20ul.
Gel Electrophoresis
- The product of digestion was run on agarose gel
- This was done in order to confirm the presence of the limonene biobrick
