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

From 2014.igem.org

(Difference between revisions)
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                         <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed identities of all parts, but not whole sequence of the parts was covered</li>
                         <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed identities of all parts, but not whole sequence of the parts was covered</li>
                         <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>kivD</i></li>
                         <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>kivD</i></li>
 +
                    </ul>
 +
          </ul>
 +
                  <li><b><i>alsS</i></b></li>
 +
          <ul>
 +
    <li>This week we aim to transform the construct from the parts distribution.</li>
 +
                    <ul>
 +
                <li>Transformation with electrocompotetent cells</li>
 +
                        <ul>
 +
                          <li>K539627: pSB1C3-<i>alsS</i> (parts distribution from 2013, plate 1, 24H)</li>
 +
                        </ul>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>alsS</i></li>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed identities of all parts, but not whole sequence of the parts was covered</li>
 +
                        <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>alsS</i></li>
 +
                    </ul>
 +
          </ul>
 +
                  <li><b><i>ilvC</i></b></li>
 +
          <ul>
 +
    <li>This week we aim to transform the construct from the parts distribution.</li>
 +
                    <ul>
 +
                <li>Transformation with electrocompotetent cells</li>
 +
                        <ul>
 +
                          <li>K539621: pSB1C3-<i>ilvC</i> (parts distribution from 2013, plate 1, 24F)</li>
 +
                        </ul>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>alsS</i></li>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed identities of all parts, but not whole sequence of the parts was covered</li>
 +
                        <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>ilvC</i></li>
 +
                    </ul>
 +
          </ul>
 +
                  <li><b><i>ilvD</i></b></li>
 +
          <ul>
 +
    <li>This week we aim to transform the construct from the parts distribution.</li>
 +
                    <ul>
 +
                <li>Transformation with electrocompotetent cells</li>
 +
                        <ul>
 +
                          <li>K539626: pSB1C3-<i>ilvD</i> (parts distribution from 2013, plate 1, 22J)</li>
 +
                        </ul>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>alsS</i></li>
 +
                        <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed identities of all parts, but not whole sequence of the parts was covered</li>
 +
                        <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>ilvD</i></li>
                     </ul>
                     </ul>
-
 
          </ul>
          </ul>
               </ul>
               </ul>

Revision as of 10:10, 27 August 2014


June

Week 1     06/02 - 06/08
Week 1     06/02 - 06/08
  • kivD
    • This week we aim to transform the construct from the parts distribution.
      • Transformation with electrocompotetent cells
        • K538742: pSB1C3-kivD (parts distribution from 2013, plate 1, 24L)
      • Plasmid isolation of kivD
      • Sequencing confirmed identities of all parts, but not whole sequence of the parts was covered
      • Glycerin stock created: E. coli KRX pSB1C3-kivD
  • alsS
    • This week we aim to transform the construct from the parts distribution.
      • Transformation with electrocompotetent cells
        • K539627: pSB1C3-alsS (parts distribution from 2013, plate 1, 24H)
      • Plasmid isolation of alsS
      • Sequencing confirmed identities of all parts, but not whole sequence of the parts was covered
      • Glycerin stock created: E. coli KRX pSB1C3-alsS
  • ilvC
    • This week we aim to transform the construct from the parts distribution.
      • Transformation with electrocompotetent cells
        • K539621: pSB1C3-ilvC (parts distribution from 2013, plate 1, 24F)
      • Plasmid isolation of alsS
      • Sequencing confirmed identities of all parts, but not whole sequence of the parts was covered
      • Glycerin stock created: E. coli KRX pSB1C3-ilvC
  • ilvD
    • This week we aim to transform the construct from the parts distribution.
      • Transformation with electrocompotetent cells
        • K539626: pSB1C3-ilvD (parts distribution from 2013, plate 1, 22J)
      • Plasmid isolation of alsS
      • Sequencing confirmed identities of all parts, but not whole sequence of the parts was covered
      • Glycerin stock created: E. coli KRX pSB1C3-ilvD
  • 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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