05/08/13

From 2013.igem.org

(Difference between revisions)
(Gel Electrophoresis)
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*The product of digestion was run on agarose gel
*The product of digestion was run on agarose gel
**This was done in order to confirm the presence of the limonene biobrick
**This was done in order to confirm the presence of the limonene biobrick
 +
[[File:iGem_Digested_050813.jpg]]

Revision as of 09:02, 6 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.
samplevolume(ul)concentration (ng/ul)yield(ug)
5.199757.4
5.210044.24.4
5.39846.74.6
10.14661.82.84
10.24477.13.39
10.34713.70.64

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
IGem Digested 050813.jpg