Team:Bielefeld-Germany/Labjournal/week20

From 2012.igem.org

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(Tuesday September 11th)
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===Tuesday September 11th===
===Tuesday September 11th===
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* '''Team Shuttle Vector:'''
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* '''Team Shuttle Vector:''' Digest of shuttle shuttle vector with ''Pvu''II and ''Hind''III as control. Agarose gel looks good.  
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Digest of shuttle shuttle vector with ''Pvu''II and ''Hind''III as control. Agarose gel looks good.  
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* '''Fungal Laccases:''' PCR on tvel5 laccase for cloning in shuttle vector. Digest of shuttle vector and digest of tvel35 with ''Aar''I enzyme.
* '''Fungal Laccases:''' PCR on tvel5 laccase for cloning in shuttle vector. Digest of shuttle vector and digest of tvel35 with ''Aar''I enzyme.

Revision as of 21:32, 19 September 2012

Contents

Week 20 (09/10 - 09/16/12)

Monday September 10th

  • Team Cultivation & Purification:
    • Cell disruption via high-pressure homogeniser and purification via Ni-NTA column were performed for the following samples: 3L fermentation of E.coli KRX with laccase from B.pumilus (09/09), 6L fermentation of E.coli KRX with laccase from E.coli (09/07) and B.pumilus (09/09).
    • We made SDS-Pages of purification of laccase from B.pumilus
  • Team Site Directed Mutagenesis:
    • Digestion of the six tvel10-plasmids. Three were unmutated and other three hadn't lost the illegal SpeI-restriction-site, but their second fragment was of a smaller size.
    • plated three additional tvel-t243g-colonies.

Tuesday September 11th

  • Team Shuttle Vector: Digest of shuttle shuttle vector with PvuII and HindIII as control. Agarose gel looks good.
  • Fungal Laccases: PCR on tvel5 laccase for cloning in shuttle vector. Digest of shuttle vector and digest of tvel35 with AarI enzyme.

Wednesday September 12th

  • Team Site Directed Mutagenesis:
    • Plasmid-isolation of the three tvel10-plasmids and digestion with SpeI showed two unmutated plasmids and one with the same wrong restriction-fragments as Monday. There must be a systematical error. pfu-PCR should be done again.

Thursday September 13th

Friday September 14th

Saturday September 15th

Sunday September 16th

Sunday


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