12/09/13

From 2013.igem.org

(Difference between revisions)
(Digestion of the samples purified with the enzyme PstI-HF)
(Digestion of the samples purified with the enzyme PstI-HF)
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[[File:doubledigest_gelpurification_12092013.jpg]]
[[File:doubledigest_gelpurification_12092013.jpg]]
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The picture above shows that the assumption of the PstI enzyme not working properly was wrong, as it shows the same result as the gel run on the
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The picture above shows that the assumption of the PstI enzyme not working properly was wrong, as it shows the same result as the gel run on the [[11/09/13]].

Revision as of 20:31, 13 September 2013

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Contents

Isolate plasmid from overnight culture

  • Using Omega mini prep kit protocol
  • Samples:
    • TODF 1,2,3,
    • TODX 1, 2, 3
    • TOBG 1, 2, 3,
    • RFP 1, 2, 3
sampleVolume (ul)Concentration (ng/ul)
TODF143120
TODF24398.8
TODX142399.3
TODX244442.2
TODX343417.5
TOBG143262.4
TOBG247319.2
TOBG341338.4
RFP14177.7
RFP24132.7
RFP34259

Glycerol stocks

  • Took 750ul from overnight culture and centrifuged at full speed for ~2 minutes
  • Supernatant was removed
  • Pellet resuspended in 375ul of HMFM
  • Samples were then frozen at -80

Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13

The ge was purified by using the The Zymoclean Gel DNA recovery Kit and its protocol was followed.

Nanodrop Results

SampleVolumeConcentration (ng/ul)280/260260/230
TODX48.57.11.830.72
TODF45.521.71.891.19
TOBG47.791.770.70
pSB1C3 (RFP)46.27.31.810.16

Digestion of the samples purified with the enzyme PstI-HF

DNA volumeVolumeCutsmart bufferEnzyme volumedH2O added to total volume of 60 ul
TODX236130
TODF22.56130.5
TOBG226131
pSB1C3 (RFP)20.56132.5

Doubledigest gelpurification 12092013.jpg

The picture above shows that the assumption of the PstI enzyme not working properly was wrong, as it shows the same result as the gel run on the 11/09/13.