12/09/13

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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
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