Team:Yale/Our Project/test-page

From 2010.igem.org

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<div style="clear: right; text-align: left; float: left; padding: .4em .9em .9em">
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<div id="left-col">
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<h5>
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our project
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</h5>
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<p>
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<ul id="proj-nav">
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<li><a href="https://2010.igem.org/Team:Yale/Our Project">introduction</a></li>
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<li><a href="https://2010.igem.org/Team:Yale/Our Project/Applications">applications</a></li>
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<li><a href="https://2010.igem.org/Team:Yale/Our Project/Methods">methods</a></li>
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<li><b><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook">lab notebook</a></b></li>
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<li id="nb"><b><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook">week 1</a></b></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 2">week 2</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 3">week 3</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 4">week 4</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 5">week 5</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 6">week 6</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 7">week 7</a></li>
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<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 8">week 8</a></li>
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<li><a href="https://2010.igem.org/Team:Yale/Our Project/Protocols">protocols</a></li>
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<li><a href="https://2010.igem.org/Team:Yale/Our Project/Safety">safety</a></li>
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<li><a href="https://2010.igem.org/Team:Yale/Our Project/Future Work">future work</a></li>
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</ul>
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</div>
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<div id="right-col">
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<h5>
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lab notebook: week 1 (6/7 - 6/13)
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</h5>
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<ul>
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<!------------- LAB NOTEBOOK ------------->
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<li>
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Monday 6/7--We successfully defend our project to our sponsors(yay!)
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</li>
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<a id="link" href="#" onclick="toggle_visibility('monday'); return false;">See more</a>
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<div style="display:none" id="monday">
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<!------------- Monday ------------->
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Extra content
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<!------------- Monday ------------->
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<h3>People</h3>
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'''[[User:Thomas Dziubla| Principal Investigator]]''' <br>
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'''Graduate Students''' {{hide|
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* [[User: David Cochran| David Cochran]]
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* [[User:John M. Medley| John M. Medley]]
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* [[User: Sundar Prasanth| Sundar Prasanth]]
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* [[User:Paritosh Wattamwar| Paritosh Wattamwar]]
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}}
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'''Undergraduates''' {{hide|
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* [[Matt Sean Alexander]]
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* [[Eric Beane]]
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* [[User:Kelly C. Cummins|Kelly Cummins]]
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}}
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'''High School''' {{hide|
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* [[David Parell]]
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}}
</div>
</div>
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<li>
 
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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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<div style="clear: right; text-align: left; float: left; padding: .4em .9em .9em">
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<h3>Research</h3>
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<a id="link" href="#" onclick="toggle_visibility('wednesday'); return false;">See more</a>
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'''[[Dziubla:Research|Research Overview]]'''
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<div style="display:none" id="wednesday">
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'''[[Dziubla:Projects|Projects]]''' {{hide|
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* [[Dziubla:Projects|Antioxidant Polymers]]
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<!------------- Wednesday------------->
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* [[Dziubla:Projects|Adhesion Barriers]]
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Began growing two strains of E. coli--LE392 chosen as a generally healthy option, and DH5alpha chosen because Keasling et. al. used it in their thiosulfate reductase work.
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'''[[Dziubla:Publications|Publications]]'''<br>
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    * Spread samples of LE392 (Grindley lab) and DH5alpha (Modis lab) on LB plates and left to incubate overnight at 37
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'''[[Dziubla:Links|Links]]'''
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<!------------- Tuesday------------->
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</div>
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<li>
 
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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 
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<a id="link" href="#" onclick="toggle_visibility('thursday'); return false;">See more</a>
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<!------------- Thursday ------------->
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<h4> Baseline Wide Concentration Range Copper Growth Assay </h4>
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Set out to find baseline growth exhibited by the E. coli strains being used when grown in a variety of copper concentrations. This will serve as a basis for comparison once copper tolerance has been engineered. Chose a range from the very, very low (sure to have no effect) to the very, very high (surely lethal).
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<h3>Resources</h3>
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Set-up
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'''[[Dziubla:Protocols|Protocols]]'''<br>
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'''[[Dziubla:Equipment|Equipment]]''' {{hide|
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    * Studies were conducted in a 96-well plate, and growth was measured with a plate reader.
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* DLS
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    * 12 concentrations of copper(II) sulfate/LB solution were used with final (post-E. coli addition) copper(II) sulfate concentrations of 1 M, 0.1 M, 50 mM, 10 mM, 1 mM, 500 uM, 250 uM, 100 uM, 10 uM, 1 uM, 0.1 uM, and 0 uM, and each concentration was given its own column in the well plate. A row of cells with just the copper solutions was included to function as blanks.
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* Glovebox
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* Fluorimeter
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layout of 96-well plate
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* Freeze Dryer
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layout of 96-well plate
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* Uv-Vis
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* GPC/HPLC
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    * Each of the two E. coli strains (LE392 and DH5alpha) was grown with each of the copper concentrations from three different starting densities. These three initial densities (OD 0.075,OD 0.0375, and OD 0.01875) were chosen so that mid-log phase would be reached at 1.5 hr, 2 hr. and 2.5 hr respectively after the start of the trial.
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    * Absorption measurements were taken approximately every 10 minutes for four hours. Lack of a 600 nm filter necessitated measurement at 540 nm, a wavelength at which copper (II) sulfate absorbs, but the use of the appropriate blanks prevented this from interfering with data collection.
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'''[[Dziubla:Materials|Materials]]'''<br>
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    * Samples were kept at 37 with alternated minutes of orbital shaking and stillness (plate reader does not allow continual shaking).
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'''[[Dziubla:Back Door|Internal]]'''<br>
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<h4> Results & Conclusions </h4>
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    * Both cells lines exhibit very similar copper tolerances, growing without noticeable effect at concentrations up to 500 uM, experiencing slowed growth at 1 mM, and no growth at 10 mM.
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    * More trials are necessary at concentration ranges between 500 uM and 10 mM to determine more exactly the maximum copper tolerance of these cell lines.
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    * After four hours of growth the healthier cells were still growing very rapidly. Future assays will extend recording time to five hours in an effort to capture the plateau region.
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<i> This work is also documented on pages 1-5 in the hard copy of the lab notebook </i>
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<!------------- Thursday------------->
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<h3>Contact</h3>
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Thomas Dziubla<br>
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Friday 6/11--Ran another growth assay with middling concentrations based on results of first, but bacteria failed to grow well--b/c allowed to overgrow?
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Assistant Professor <br>
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Mat and Chem Engineering <br>
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</li>
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University of Kentucky<br>
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<a id="link" href="#" onclick="toggle_visibility('friday'); return false;">See more</a>
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177 F Paul Anderson Tower<br>
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<div style="display:none" id="friday">
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Lexington, KY 40506-0046<br>
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<!------------- Friday ------------->
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dziubla-at-engr.uky.edu<br>
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Extra content
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<!------------- Friday ------------->
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__NOTOC__
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<!------------- LAB NOTEBOOK ------------->
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[[Category:Dziubla]]
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Revision as of 06:13, 25 October 2010

850px

People

Principal Investigator
Graduate Students

Template loop detected: Template:Hide Undergraduates

Template loop detected: Template:Hide High School

Template loop detected: Template:Hide

Resources

Protocols
Equipment

  • DLS
  • Glovebox
  • Fluorimeter
  • Freeze Dryer
  • Uv-Vis
  • GPC/HPLC

Template loop detected: Template:Hide Materials
Internal

Contact

Thomas Dziubla
Assistant Professor
Mat and Chem Engineering
University of Kentucky
177 F Paul Anderson Tower
Lexington, KY 40506-0046
dziubla-at-engr.uky.edu

Retrieved from "http://2010.igem.org/Team:Yale/Our_Project/test-page"