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

From 2014.igem.org

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<ul>
<ul>
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<li>Transformation with electrocompotetent <i> E.coli </i> KRX cells:</li>
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<li>Transformation of electrocompotetent <i> E.coli </i> KRX cells:</li>
<ul>
<ul>
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<li>K538742: <i>kivD</i> (parts distribution from 2013, plate 1, 24L)</li>
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<li>K538742: <i>pSB1C3-kivD</i> (parts distribution from 2013, plate 1, 24L)</li>
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<li>K539621: <i>ilvC</i> (parts distribution from 2013, plate 1, 24F)</li>
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<li>K539621: <i>pSB1C3-ilvC</i> (parts distribution from 2013, plate 1, 24F)</li>
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<li>K539626: <i>ilvD</i> (parts distribution from 2013, plate 1, 22J)</li>
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<li>K539626: <i>pSB1C3-ilvD</i> (parts distribution from 2013, plate 1, 22J)</li>
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<li>K539627: <i>alsS</i> (parts distribution from 2013, plate 1, 24H)</li>
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<li>K539627: <i>pSB1C3-alsS</i> (parts distribution from 2013, plate 1, 24H)</li>
</ul>
</ul>
<li> Resulting colonies were selected to start liquid cultures: </li>
<li> Resulting colonies were selected to start liquid cultures: </li>
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<li>Plasmid isolation by ThermoScientifici MiniPrep</li>
<li>Plasmid isolation by ThermoScientifici MiniPrep</li>
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<li>Sanger sequencing using <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VF" target="_blank">VF</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VR" target="_blank">VR</a> as sequencing primers</li>
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<li>Sanger sequencing using <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VF-Primer" target="_blank">VF</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#VR-Primer" target="_blank">VR</a> as sequencing primers</li>
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<ul>
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<li>Identities of all parts were confirmed, but not whole sequence of the parts was covered</li>
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</ul>
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<li>Glycerin stocks were created for all four strains:</li>
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<ul>
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<li><i>E. coli</i> KRX <i>pSB1C3-alsS</i></li>
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<li><i>E. coli</i> KRX <i>pSB1C3-ilvC</i></li>
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<li><i>E. coli</i> KRX <i>pSB1C3-ilvD</i></li>
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<li><i>E. coli</i> KRX <i>pSB1C3-kivD</i></li>
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</ul>
</ul>
</ul>
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Revision as of 04:58, 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 ThermoScientifici 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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