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05/08/13
From 2013.igem.org
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| + | {| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center" | ||
| + | !align="center"|[[Team:Leicester|Home]] | ||
| + | !align="center"|[[Team:Leicester/Team|Team]] | ||
| + | !align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile] | ||
| + | !align="center"|[[Team:Leicester/Project|Project]] | ||
| + | !align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]] | ||
| + | !align="center"|[[Team:Leicester/Modeling|Modeling]] | ||
| + | !align="center"|[[Team:Leicester/Notebook|Notebook]] | ||
| + | !align="center"|[[Team:Leicester/Safety|Safety]] | ||
| + | !align="center"|[[Team:Leicester/Attributions|Attributions]] | ||
| + | |} | ||
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==Isolation of ligated plasmid== | ==Isolation of ligated plasmid== | ||
*The ligated plasmid- chloramphenicol backbone and limonene biobrick, were isolated from cells that were incubated overnight in broth by miniprep. | *The ligated plasmid- chloramphenicol backbone and limonene biobrick, were isolated from cells that were incubated overnight in broth by miniprep. | ||
Revision as of 14:14, 5 August 2013
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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
