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05/08/13
From 2013.igem.org
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**varying concentration of dH2O and plasmid depending on overall concentration of plasmid in each tube. | **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. | ** 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 | ||
Revision as of 14:02, 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.
- 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
