Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun

From 2014.igem.org

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<li>Products were analyzed by agarose gel electrophoresis</li>
<li>Products were analyzed by agarose gel electrophoresis</li>
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<li> PCR conditions need to be optimized do to low product quality and missing products</li>
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<li> PCR conditions need to be optimized due to low product quality and missing products</li>
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Revision as of 05:31, 27 August 2014


June

  • Transformation of electrocompotetent E.coli KRX cells:
    • K538742: pSB1C3-kivD (parts distribution from 2013, plate 1, 24L)
    • K539621: pSB1C3-ilvC (parts distribution from 2013, plate 1, 24F)
    • K539626: pSB1C3-ilvD (parts distribution from 2013, plate 1, 22J)
    • K539627: pSB1C3-alsS (parts distribution from 2013, plate 1, 24H)
  • Resulting colonies were selected to start liquid cultures:
    • 5 ml of LB in 20 ml reaction tube
    • incubation over night at 37°C and 250 rpm
  • Plasmid isolation by ThermoScientific MiniPrep
  • Sanger sequencing using VF and VR as sequencing primers
    • Identities of all parts were confirmed, but not whole sequence of the parts was covered
  • Glycerin stocks were created for all four strains:
    • E. coli KRX pSB1C3-alsS
    • E. coli KRX pSB1C3-ilvC
    • E. coli KRX pSB1C3-ilvD
    • E. coli KRX pSB1C3-kivD

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