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Team:Bielefeld-Germany/Labjournal/week20
From 2012.igem.org
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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