Difference between revisions of "Team:Rotterdam HR/Results"

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<div class="text-container">
 
<div class="text-container">
 
<h1>General</h1>
 
<h1>General</h1>
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<p style="text-align: left !important;"><img src="https://static.igem.org/mediawiki/parts/5/5e/T--Rotterdam_HR--foto1Res.jpeg" style="width: 360px; height: 300px; float: right;">
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<p style="text-align: left !important;"><img src="https://static.igem.org/mediawiki/parts/5/5e/T--Rotterdam_HR--foto1Res.jpeg"
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style="width: 360px; height: 300px; float: right;">
 
Our plan was to create multiple composite parts, which included CooA.
 
Our plan was to create multiple composite parts, which included CooA.
−
To be pure the DNA needed to have a A260/280 value between 1,80-2,00 and a A260/230 value between 2,00 and 2,20. These composite parts would have different
+
To be pure the DNA needed to have a A260/280 value between 1,80-2,00 and a A260/230 value between 2,00 and 2,20.
 +
These composite parts would have different
 
promoters and output genes, so we can measure carbon monoxide in multiple ways.
 
promoters and output genes, so we can measure carbon monoxide in multiple ways.
 
</p>
 
</p>
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Hereby there would be looked at the curve of the line also. If the line wasn’t smooth, the DNA was most likely not
 
Hereby there would be looked at the curve of the line also. If the line wasn’t smooth, the DNA was most likely not
 
pure. A pure sample has a curve like shown in figure 1.
 
pure. A pure sample has a curve like shown in figure 1.
 +
</p>
 +
<p>
 +
To produce enough DNA of BBa_J04450 (an plasmid containing a functional spacer) we performed multiple mini preps (DNA
 +
isolations), which gave us the following results:
 +
</p>
 +
<table>
 +
<tbody>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 1</td>
 +
<td>270,71 ng/µL</td>
 +
</tr>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 2</td>
 +
<td>119,84 ng/µL</td>
 +
</tr>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 1</td>
 +
<td>451,17 ng/µL</td>
 +
</tr>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 2</td>
 +
<td>319,01 ng/µL</td>
 +
</tr>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 1</td>
 +
<td>136,98 ng/µL</td>
 +
</tr>
 +
<tr>
 +
<td><a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a></td>
 +
<td><a href="http://parts.igem.org/Part:pSB1C3">pSB1C3</a> 2</td>
 +
<td>80,90 ng/µL</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<p>
 +
Successful isolated DNA could be digested and ligated with another biobrick for testing. Early named biobricks were
 +
paired with each other, where BBa_K133071 and BBa_K173003 functioned as vectors and BBa_I134353 and BBa_J23100 as
 +
inserts. We used the backbone from the vectors, so we only needed to insert 1 biobrick into another biobrick. To know
 +
if the ligation had worked the DNA was transformed into the E.coli strain: NEB10Bèta. Colonies that grow were tested
 +
by digesting again. The digested DNA was put on gel, and the length was determined.
 
</p>
 
</p>
 
</div>
 
</div>

Revision as of 00:13, 18 October 2018

Results

Results

General

Our plan was to create multiple composite parts, which included CooA. To be pure the DNA needed to have a A260/280 value between 1,80-2,00 and a A260/230 value between 2,00 and 2,20. These composite parts would have different promoters and output genes, so we can measure carbon monoxide in multiple ways.

For further steps we isolated the DNA. The DNA had to be pure for digestions. To be sure the DNA was pure, requirements were set. Hereby there would be looked at the curve of the line also. If the line wasn’t smooth, the DNA was most likely not pure. A pure sample has a curve like shown in figure 1.

To produce enough DNA of BBa_J04450 (an plasmid containing a functional spacer) we performed multiple mini preps (DNA isolations), which gave us the following results:

BBa_J04450 pSB1C3 1 270,71 ng/µL
BBa_J04450 pSB1C3 2 119,84 ng/µL
BBa_J04450 pSB1C3 1 451,17 ng/µL
BBa_J04450 pSB1C3 2 319,01 ng/µL
BBa_J04450 pSB1C3 1 136,98 ng/µL
BBa_J04450 pSB1C3 2 80,90 ng/µL

Successful isolated DNA could be digested and ligated with another biobrick for testing. Early named biobricks were paired with each other, where BBa_K133071 and BBa_K173003 functioned as vectors and BBa_I134353 and BBa_J23100 as inserts. We used the backbone from the vectors, so we only needed to insert 1 biobrick into another biobrick. To know if the ligation had worked the DNA was transformed into the E.coli strain: NEB10Bèta. Colonies that grow were tested by digesting again. The digested DNA was put on gel, and the length was determined.