Team:Davidson-MissouriW/Notebook

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<li>IPTG experiment started using varying tetracycline concentrations.
<li>IPTG experiment started using varying tetracycline concentrations.
<li>Cell count data collected on IPTG TetA experiment. </li>
<li>Cell count data collected on IPTG TetA experiment. </li>
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</ul>
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<br>
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<br>'''July 23, 2010'''
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<hr>
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<ul>
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<li>Cell count data collected from IPTG experiment..
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<li>Flourimeter data collected from IPTG experiment. </li>
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</ul>
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<br>
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<br>'''July 26, 2010'''
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<hr>
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<ul>
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<li>Presented Missouri Western data to Davidson colleagues.
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<li>Presented Davidson data to Missouri Western colleagues. </li>
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</ul>
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<br>
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<br>'''July 27, 2010'''
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<hr>
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<ul>
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<li>Worked on team wiki. </li>
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</ul>
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<br>
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<br>'''July 28, 2010'''
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<hr>
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<ul>
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<li>n/a
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<li>n/a </li>
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</ul>
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<br>
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<br>'''July 30, 2010'''
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<hr>
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<ul>
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<li>n/a
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<li>n/a </li>
</ul>
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Revision as of 14:53, 27 July 2010

Notebook

May 31, 2010

  • Tested to find lethal concentration of Tet in plated and liquid mediums


June 2, 2010

  • Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP


June 3, 2010

  • Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP


June 4, 2010

  • Mini prepped RBS-TetA-TT
  • RX Fragment digestion for RBS-TetA-TT


June 7, 2010

  • Ligate and plasmid check for pSB1A2 or pSB1AK3


June 8, 2010

  • Double digest on pBAD-RBS-RFP to make an RS fragment insert.


June 9, 2010

  • EcoRI and PstI digest on pLac and pBad colonies.
  • Arabinose and IPTG concentration experiments started.


June 10, 2010

  • Picked 14 colonies from pLac-RBS-RFP and pLac-RBS-RFP-RBS plates.
  • Started inducer curve experiments.


June 11, 2010

  • Ligated S03736 into pSB1A7 vector.
  • EcoRI/PstI digest on pLac-RBS-RFP-RBS colonies.


June 14, 2010

  • EcoRI/PstI diagnostic digest performed on pBad-RBS-RFP colonies.
  • Inducer curve experiment started on pBad and pLac constructs.


June 15, 2010

  • Ligated pLac-RBS-RFP + RBS-TetA.
  • IPTG experiment started on pLacI-RBS-RFP


June 16, 2010

  • EcoRI/PstI diagnostic digest performed on pLac-RBS-RFP in pSB1A7 vector.
  • IPTG experiment started on plates with pLac and pLacI constructs.


June 17, 2010

  • Flourimeter data collected from IPTG experiment.
  • Ligated pLacI-RBS-RFP + RBS-TetA.


June 18, 2010

  • Picked 4 colonies from pLacI-RBS-RFP-RBS-TetA plates.
  • IPTG experiment started using newly created IPTG.


June 21, 2010

  • Digested TetA with EcoRI/NheI.
  • Annealed oligos for optimized and deoptimized TetA segment 1.


June 22, 2010

  • Ligated annealed TetA oligos into digested TetA.
  • Inducer curve experiment started on pLac and pLacI constructs.


June 23, 2010

  • Preparative NheI/BamHI digest performed on TetA.
  • Preparative EcoRI/NheI digest performed on TetA.


June 24, 2010

  • Annealed optimized and deoptimized segment 1 TetA ligated into vector.
  • Annealed optimized and deoptimized segment 2 TetA ligatied into vector.


June 27, 2010

  • Inducer curve experiment started on pLac and pLacI constructs.
  • Picked colonies from TetA segment 1 & 2 optimized & deoptimized plates.


June 29, 2010

  • Screened segment 1 optimized and deoptimized candidate clones using RFLP
  • Screened segment 2 optimized and deoptimized candidate clones using RFLP


June 30, 2010

  • Tet titration experiment performed on pLac and pLacI +RBS-RFP-RBS-TetA.
  • Ligated TetA onto pLac and pLacI +RBS-RFP-RBS-TetA with no terminator.


July 1, 2010'

  • Preparative BamHI/NheI digest performed on TetA.
  • New experiment protocol developed for reduced vector background noise.


July 2, 2010

  • Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP


July 6, 2010

  • Ligated S04446 and S04447 to J31007.
  • Tet titration experiment performed on pLac and pLacI +RBS-RFP-RBS-TetA.


July 7, 2010

  • Ligated S04448 to RBS-TetA.
  • Ligated S04449 to RBS-TetA.


July 8, 2010

  • IPTG experiment started using pre-induced method.
  • IPTG experiment started using varying levels of tetracycline.


July 9, 2010

  • Picked optimized and deoptimized TetA colonies.
  • Flourimeter data collected from IPTG experiment.


July 10, 2010

  • Used RFLP to screen for candidate TetA clones.
  • Transformed various pLac and pLacI constructs into MG1655 cells.


July 13, 2010

  • Used RFLP to screen for candidate TetA clones.
  • EcoRI/PstI diagnostic digest performed on K199150.


July 14, 2010

  • Ligated segment 2 optimized and deoptimzed TetA behind segment 1.
  • IPTG experiment started on I715039-1 and I715039-2.


July 15, 2010

  • Cell count data collected from IPTG experiments.
  • Flourimeter data collected from IPTG experiments.


July 16, 2010

  • Tet titration experiment started using pSB3T5 vector.
  • Tet titration experiment started using optimized and deoptimized TetA.


July 19, 2010

  • Cell count data collected from Tet titrations.
  • Tet titration experiment started using optimized and deoptimized TetA.


June 3, 2010

  • Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP


July 20, 2010

  • Flourimeter data collected on IPTG experiment.


July 21, 2010

  • IPTG experiment started on optimized and deoptmized TetA constructs.
  • Passage I715039-1 and I715039-2 experiments started.


July 22, 2010

  • IPTG experiment started using varying tetracycline concentrations.
  • Cell count data collected on IPTG TetA experiment.



July 23, 2010

  • Cell count data collected from IPTG experiment..
  • Flourimeter data collected from IPTG experiment.



July 26, 2010

  • Presented Missouri Western data to Davidson colleagues.
  • Presented Davidson data to Missouri Western colleagues.



July 27, 2010

  • Worked on team wiki.



July 28, 2010

  • n/a
  • n/a



July 30, 2010

  • n/a
  • n/a


Retrieved from "http://2010.igem.org/Team:Davidson-MissouriW/Notebook"