Team:UCL/Science/Experiment

From 2014.igem.org

(Difference between revisions)
Line 101: Line 101:
<p>Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks</p>
<p>Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks</p>
<hr>
<hr>
 +
<a name="Expt03"><h4>Experiment 3: Diagnostic digest of azo-reductase plasmids - Date: [10/07/2014]</h4></a>
<a name="Expt03"><h4>Experiment 3: Diagnostic digest of azo-reductase plasmids - Date: [10/07/2014]</h4></a>
 +
<p><strong>Protocols</strong><br/>  
<p><strong>Protocols</strong><br/>  
<a href="/Team:UCL/protocols"><span class="label-warning">competent cells</span></a>
<a href="/Team:UCL/protocols"><span class="label-warning">competent cells</span></a>
Line 108: Line 110:
<a href="/Team:UCL/protocols"><span class="label-warning">digest</span></a>
<a href="/Team:UCL/protocols"><span class="label-warning">digest</span></a>
<a href="/Team:UCL/protocols"><span class="label-warning">gel</span></a></p>
<a href="/Team:UCL/protocols"><span class="label-warning">gel</span></a></p>
-
<p>senectus et netus et malesuada</p>
+
<h4>Background</h4>
 +
<p>Set up double digest as followed CHECK ENZYMES</p>
 +
<table style="width: 624px;">
 +
<tbody>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>Lane</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>Gene</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>DNA conc</p>
 +
 
 +
<p>(ng/l)</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>*Volume (ul)</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>H2O (ul)</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>1</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>AzoR</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>48</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>5.21</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>1.79</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>2</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>AzoR1B6</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>68</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>3.67</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>3.33</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>3</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>CotA</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>103</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>2.42</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>4.58</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>4</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>BsDyP</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>51</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>4.91</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>2.09</p>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<p>5</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>PyDyp</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>55</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>4.54</p>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<p>2.46</p>
 +
</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<p>Performed transformation of the following BioBricks using non-commercial competent cells:</p>
 +
<br>
 +
<ol>
 +
<li>BBa_J23119</li>
 +
<li>BBa_R0010</li>
 +
<li>BBa_R0011</li>
 +
<li>BBa_K206000</li>
 +
<li>BBa_B0034</li>
 +
<li>BBa_B0012</li>
 +
<li>BBa_J04450</li>
 +
<li>BBa_E2050</li>
 +
</ol>
 +
<p>Performed transformations of Lisbon plasmids using NEB competent cells (c2987):</p>
 +
<ol>
 +
<li>pPAzoR - P. putida Azo reductase</li>
 +
<li>pCotA - Laccase</li>
 +
<li>pPpDyP - P.Putida dye decolourizing peroxidase</li>
 +
<li>pBsDyP - B. subtilis dye decolourizing peroxidase</li>
 +
<li>p1B6 - Azoreductase 1B6 (mutated from AzoR)</li>
 +
</ol>
 +
<h4>Results</h4>
 +
<br>
 +
<p>Lisbon plasmid transformation using NEB competent cells successfully grew when plated with the appropriate antibiotic</p>
 +
<p>BioBrick plasmid transformations were successful, however the colonies had grown as a lawn making it difficult to pick a single colony.</p>
 +
<br>
 +
 +
<h4>Conclusion</h4>
 +
<p>Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks</p>
 +
<hr>
-
<a name="Expt04"><h4>Experiment 4: Creation of azo-reductase BioBrick parts from plasmids - Date: [ddmmyy]</h4></a>
+
<a name="Expt04"><h4>Experiment 4: Creation of azo-reductase BioBrick parts from plasmids - Date: [17/07/14]</h4></a>
<p><strong>Protocols</strong><br/>  
<p><strong>Protocols</strong><br/>  
<a href="/Team:UCL/protocols"><span class="label-warning">competent cells</span></a>
<a href="/Team:UCL/protocols"><span class="label-warning">competent cells</span></a>

Revision as of 17:09, 8 September 2014

Goodbye Azodye UCL iGEM 2014

Experiments

Laboratory Team

List of Experiments

  1. Experiment 01: Extraction of useful BioBrick plasmids from iGEM 2014 Distribution Kit
  2. Experiment 02: Transforming E. coli with Azo-reductase plasmids
  3. Experiment 03: Diagnostic digest of azo-reductase plasmids
  4. Experiment 04: Creation of azo-reductase BioBrick parts from plasmids
  5. Experiment 05: Diagnostic digest of azo-reductase BioBrick parts
  6. Experiment 06: Assembling azo-reductase BioBrick Device(s)
  7. Experiment 07: Characterisation of azo-reductase BioBrick devices

Experiment 1: Extraction of useful BioBrick plasmids from iGEM 2014 Distribution Kit - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

Background

Content

Results

Content Here

Conclusion

Content Here

Experiment 2: Transforming E. coli with Azo-reductase plasmids - Date: [10/07/2014]

Protocols
competent cells transformation miniprep digest gel


Background


Performed transformation of the following BioBricks using non-commercial competent cells:


  1. BBa_J23119
  2. BBa_R0010
  3. BBa_R0011
  4. BBa_K206000
  5. BBa_B0034
  6. BBa_B0012
  7. BBa_J04450
  8. BBa_E2050

Performed transformations of Lisbon plasmids using NEB competent cells (c2987):

  1. pPAzoR - P. putida Azo reductase
  2. pCotA - Laccase
  3. pPpDyP - P.Putida dye decolourizing peroxidase
  4. pBsDyP - B. subtilis dye decolourizing peroxidase
  5. p1B6 - Azoreductase 1B6 (mutated from AzoR)

Results


Lisbon plasmid transformation using NEB competent cells successfully grew when plated with the appropriate antibiotic

BioBrick plasmid transformations were successful, however the colonies had grown as a lawn making it difficult to pick a single colony.


Conclusion

Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks

Experiment 3: Diagnostic digest of azo-reductase plasmids - Date: [10/07/2014]

Protocols
competent cells transformation miniprep digest gel

Background

Set up double digest as followed CHECK ENZYMES

Lane

Gene

DNA conc

(ng/l)

*Volume (ul)

H2O (ul)

1

AzoR

48

5.21

1.79

2

AzoR1B6

68

3.67

3.33

3

CotA

103

2.42

4.58

4

BsDyP

51

4.91

2.09

5

PyDyp

55

4.54

2.46

Performed transformation of the following BioBricks using non-commercial competent cells:


  1. BBa_J23119
  2. BBa_R0010
  3. BBa_R0011
  4. BBa_K206000
  5. BBa_B0034
  6. BBa_B0012
  7. BBa_J04450
  8. BBa_E2050

Performed transformations of Lisbon plasmids using NEB competent cells (c2987):

  1. pPAzoR - P. putida Azo reductase
  2. pCotA - Laccase
  3. pPpDyP - P.Putida dye decolourizing peroxidase
  4. pBsDyP - B. subtilis dye decolourizing peroxidase
  5. p1B6 - Azoreductase 1B6 (mutated from AzoR)

Results


Lisbon plasmid transformation using NEB competent cells successfully grew when plated with the appropriate antibiotic

BioBrick plasmid transformations were successful, however the colonies had grown as a lawn making it difficult to pick a single colony.


Conclusion

Made overnight culture of lisbon plasmids, preparing to keep in glycerol stocks

Experiment 4: Creation of azo-reductase BioBrick parts from plasmids - Date: [17/07/14]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 5: Diagnostic digest of azo-reductase BioBrick parts - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 6: Assembling azo-reductase BioBrick Device(s) - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Experiment 7: Characterisation of azo-reductase BioBrick devices - Date: [ddmmyy]

Protocols
competent cells transformation miniprep digest gel

senectus et netus et malesuada

Contact Us

University College London
Gower Street - London
WC1E 6BT
Biochemical Engineering Department
Phone: +44 (0)20 7679 2000
Email: ucligem2014@gmail.com

Follow Us

Retrieved from "http://2014.igem.org/Team:UCL/Science/Experiment"