Team:Yale/Our Project/Notebook/Week 2

From 2010.igem.org

(Difference between revisions)
m
 
(19 intermediate revisions not shown)
Line 22: Line 22:
<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 7">week 7</a></li>
<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 7">week 7</a></li>
<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 8">week 8</a></li>
<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 8">week 8</a></li>
 +
<li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 9">week 9</a></li>
<li><a href="https://2010.igem.org/Team:Yale/Our Project/Protocols">protocols</a></li>
<li><a href="https://2010.igem.org/Team:Yale/Our Project/Protocols">protocols</a></li>
<li><a href="https://2010.igem.org/Team:Yale/Our Project/Safety">safety</a></li>
<li><a href="https://2010.igem.org/Team:Yale/Our Project/Safety">safety</a></li>
Line 47: Line 48:
<!------------- Monday ------------->
<!------------- Monday ------------->
<h4> Copper Growth Assay Work Continues </h4>
<h4> Copper Growth Assay Work Continues </h4>
 +
<div align="center">
 +
<img src="https://static.igem.org/mediawiki/2010/8/8f/Yale-growthfail.jpg" />
 +
</div>
<ul>
<ul>
<li>As the growth assay of <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook">6/11</a> showed uniformly poor growth even at the lowest copper concentrations (see Dh5alpha growth above), redid it with care not to let the liquid cultures overgrow, a possible source of the cultures' previous poor performance. (add plot)</li>
<li>As the growth assay of <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook">6/11</a> showed uniformly poor growth even at the lowest copper concentrations (see Dh5alpha growth above), redid it with care not to let the liquid cultures overgrow, a possible source of the cultures' previous poor performance. (add plot)</li>
Line 67: Line 71:
<!------------- Tuesday ------------->
<!------------- Tuesday ------------->
<h4> Results and Conclusions of Narrow Concentration Range Growth Assay of 6/14</h4>
<h4> Results and Conclusions of Narrow Concentration Range Growth Assay of 6/14</h4>
-
(Show graph)
+
<div align="center">
 +
<img src="https://static.igem.org/mediawiki/2010/d/d1/Yale-dh5alpha-narrow.jpg" />
 +
</div>
<ul>
<ul>
<li> Negative readings for the 4 mM samples suggest that there was some sort of irregularity with the blank solution, so will disregard those results. </li>
<li> Negative readings for the 4 mM samples suggest that there was some sort of irregularity with the blank solution, so will disregard those results. </li>
Line 125: Line 131:
</table>  
</table>  
*This plan was eventually altered, as promoter choices were altered and no light-inducible promoter was found. <br/>
*This plan was eventually altered, as promoter choices were altered and no light-inducible promoter was found. <br/>
-
 
+
<br />
 +
<div id="right">
 +
<img src="https://static.igem.org/mediawiki/2010/7/71/Yale-biobrick.jpg" />
 +
</div>
To make these plasmids, we will rely on the standard iGEM assembly protocol involving restriction enzymes EcoRI, XbaI, PstI, and SpeI shown in the diagram at right, but in the first ligation the B0015 terminator will take the place of C0010 and the phsABC  gene in pSB74 will take the place of B0034. <br/>
To make these plasmids, we will rely on the standard iGEM assembly protocol involving restriction enzymes EcoRI, XbaI, PstI, and SpeI shown in the diagram at right, but in the first ligation the B0015 terminator will take the place of C0010 and the phsABC  gene in pSB74 will take the place of B0034. <br/>
Line 299: Line 308:
<li>
<li>
-
Saturday 6/19--in which it is revealed there was a transformation-fail
+
Saturday 6/19--Redo of Biobrick transformations & analysis of BL21 copper growth assay
</li>
</li>
Line 309: Line 318:
<b>BL21 copper growth assay analysis</b> <br/>
<b>BL21 copper growth assay analysis</b> <br/>
Retrieved the following data regarding BL21's growth in copper solution: <br/>
Retrieved the following data regarding BL21's growth in copper solution: <br/>
-
 
+
<div align="center">
-
It appears that BL21 is slightly more sensitive to copper than the other two strains, since 3 mM levels of copper(II) sulfate are enough to almost completely inhibit BL21's growth.
+
<img src="https://static.igem.org/mediawiki/2010/f/f6/Yale-bl21.jpg" />
 +
</div>
 +
It appears that BL21 is slightly more sensitive to copper than the other two strains, since 3 mM levels of copper(II) sulfate are enough to almost completely inhibit BL21's growth. <br/>
 +
<i> The activities of this day are also recorded on page 15 of the hard copy lab notebook </i>
<!------------- Saturday ------------->
<!------------- Saturday ------------->
</div>
</div>
Line 317: Line 329:
<li>
<li>
-
Sunday 6/20--in evening inoculate 5 mL liquid cultures
+
Sunday 6/20--Observed growth of  all transformants from 6/19, so used them and the culture containing pSB74 to inoculate 5 mL liquid cultures in LB with ampicillin.  Left to grow overnight on shaker at 37˚C for miniprep the following morning. <br/>
 +
<i> The activities of this day are also recorded on page 16 of the hard copy lab notebook </i>
</li>
</li>
-
<a id="link" href="javascript:ReverseDisplay('sunday')">See more/less</a>
+
 
-
<div style="display:none;" id="sunday">
+
-
<!------------- Sunday ------------->
+
-
Extra content
+
-
<!------------- Sunday ------------->
+
-
</div>
+
<!------------- LAB NOTEBOOK ------------->
<!------------- LAB NOTEBOOK ------------->

Latest revision as of 02:49, 28 October 2010

iGEM Yale

lab notebook: week 2 (6/14-6/20)

  • Monday 6/14--Redo of 6/11 assay of bacterial growth within a narrow copper(II) concentrations after analysis of data showed uniformly poor growth.
  • See more/less
  • Tuesday 6/15--Results and analysis of the 6/14 narrow concentration range growth assay as well as planning for the creation of a standard iGEM plasmid bearing the thiosulfate reductase operon (phsABC).
  • See more/less
  • Wednesday 6/16--Checked on spotted cell survival assay, collected MOPS minimal media materials & started making component solutions
  • See more/less
  • Thursday 6/17--more minimal media work and meeting, started BL21 culture
  • See more/less
  • Friday 6/18--Arrival of plasmid pSB74, transformation of Biobricks, & copper growth assays for BL21 strain.
  • See more/less
  • Saturday 6/19--Redo of Biobrick transformations & analysis of BL21 copper growth assay
  • See more/less
  • Sunday 6/20--Observed growth of all transformants from 6/19, so used them and the culture containing pSB74 to inoculate 5 mL liquid cultures in LB with ampicillin. Left to grow overnight on shaker at 37˚C for miniprep the following morning.
    The activities of this day are also recorded on page 16 of the hard copy lab notebook