Team:Yale/Our Project/Notebook

From 2010.igem.org

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<a name="6_9"> Wednesday 6/9 </a>  --First day in lab!--got set up & started cultures of two <i> E. coli </i> strains
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Wednesday 6/9 --First day in lab!--got set up & started cultures of two <i> E. coli </i> strains
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<h4> Further Copper Growth Assays—Narrow Concentration Range </h4>
<h4> Further Copper Growth Assays—Narrow Concentration Range </h4>
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Plan to do more assays of bacterial growth in the presence of copper to determine the exact threshold at which growth is affected.  Will repeat yesterday’s assay (link) with a more targeted concentration range and a longer growth time.  
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Plan to do more assays of bacterial growth in the presence of copper to determine the exact threshold at which growth is affected.  Will repeat yesterday’s assay with a more targeted concentration range and a longer growth time.  
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<li>Once again inoculated liquid cultures of DH5alpha and LE392 in 5 mL LB using bacteria from plates established on  <a href=“#6_9”> 6/9 </a> and put on shaker to grow at 37˚C </li>
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<li>Once again inoculated liquid cultures of DH5alpha and LE392 in 5 mL LB using bacteria from plates established on  6/9 and put on shaker to grow at 37˚C </li>
<li>Let grow for approximately 6 hours, by which point the DH5alpha have reached an OD (measured at 600 nm) of 0.915and the LE392 have reached OD 1.420. </li>
<li>Let grow for approximately 6 hours, by which point the DH5alpha have reached an OD (measured at 600 nm) of 0.915and the LE392 have reached OD 1.420. </li>
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<li>Diluted them to the three initial ODs chosen on 6/10 (link) in a variety of copper (II) sulfate concentrations chosen to give more detailed data than the wide concentration range trial. The concentrations chosen were   500 μM, 600 μM, 700 μM, 800 μM, 900 μM, 1 mM, 1.25 mM, 1.5 mM, 2 mM, 2.5 mM, 3 mM, 4 mM </li>
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<li>Diluted them to the three initial ODs chosen on 6/10 in a variety of copper (II) sulfate concentrations chosen to give more detailed data than the wide concentration range trial. The concentrations chosen were 500 μM, 600 μM, 700 μM, 800 μM, 900 μM, 1 mM, 1.25 mM, 1.5 mM, 2 mM, 2.5 mM, 3 mM, 4 mM </li>
<li>Using these concentrations, ran the copper growth assay overnight using the same protocol as the previous day (link), except that the plate reader took readings for five hours instead of four to better capture all of the growth phase behavior.</li>
<li>Using these concentrations, ran the copper growth assay overnight using the same protocol as the previous day (link), except that the plate reader took readings for five hours instead of four to better capture all of the growth phase behavior.</li>
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Revision as of 01:23, 27 October 2010

iGEM Yale

lab notebook: week 1 (6/7 - 6/13)

  • Monday 6/7--We successfully defend our project to our sponsors(yay!)
  • Wednesday 6/9 --First day in lab!--got set up & started cultures of two E. coli strains
  • See more/less
  • Thursday 6/10--inoculated liquid cultures in AM, made up different copper sulfate & LB solutions, then ran an assay measuring bacterial growth in the presence of copper, trying both strains at three different initial ODs and measuring for four hours
  • See more/less
  • Friday 6/11--Ran another growth assay with middling concentrations based on results of first, but bacteria failed to grow well--perhaps because allowed to overgrow prior to assay?
  • See more/less