05/08/13

From 2013.igem.org

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* The plasmid concentration in each of the 6 tubes were measured using nanodrop.
* 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.
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{|border=1
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|sample||volume(ul)||concentration (ng/ul)||yield(ug)
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|5.1||99||75||7.4
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|5.2||100||44.2||4.4
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|5.3||98||46.7||4.6
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|-
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|10.1||46||61.8||2.84
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|10.2||44||77.1||3.39
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|-
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|10.3||47||13.7||0.64
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|}
   
   
==Digestion of ligated plasmid==
==Digestion of ligated plasmid==

Revision as of 14:29, 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.
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

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