Team:UCL/Labbook/Week16

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<p class="body_text"> <a href="https://2013.igem.org/Team:UCL/LabBook/Week1">Week 1</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week2"> Week 2</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week3"> Week 3</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week4"> Week 4</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week5"> Week 5</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week6"> Week 6</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week7"> Week 7</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week8"> Week 8</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week9"> Week 9</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week10"> Week 10</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week11"> Week 11</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week12"> Week 12</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week13"> Week 13</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week14"> Week 14</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week15"> Week 15</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week16"> Week 16</a>
<p class="body_text"> <a href="https://2013.igem.org/Team:UCL/LabBook/Week1">Week 1</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week2"> Week 2</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week3"> Week 3</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week4"> Week 4</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week5"> Week 5</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week6"> Week 6</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week7"> Week 7</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week8"> Week 8</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week9"> Week 9</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week10"> Week 10</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week11"> Week 11</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week12"> Week 12</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week13"> Week 13</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week14"> Week 14</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week15"> Week 15</a> | <a href="https://2013.igem.org/Team:UCL/Labbook/Week16"> Week 16</a>
</p>  
</p>  
 +
</div>
 +
 +
<p class="minor_title">Week 16</p>
 +
<div class="full_row">
 +
<div class="gap">
 +
</div>
 +
<p class="body_text">
 +
<b>Bacterial Labs</b>
 +
</p>
 +
<p class="body_text">
 +
Results of the inoculations of transformations of 13/09 in 4xcmp: all showed growth apart from falcons 5.3, 5.2, 1.14 and 1.2. Glycerol stocks of the rest of 19 inoculations were made.
 +
</p>
 +
<p class="body_text">
 +
Minipreps of the above inoculations were made only for 1.3, 1.7, 1.9, 1.10, 1.11, 1.15, 1.20 and 5.1 due to lack of chromatographic columns.
 +
</p>
 +
<p class="body_text">
 +
Nanodrop result of the above minipreps showed concentration values below 16.0 ng/ul.
 +
Analytical digest of miniprep samples 1-9 (prepared the day before) as well as of 5.1 and 1.15 (which showed concentrations of about 15 ng/ul) with E and P
 +
</p>
 +
 +
<p class="body_text">
 +
<table>
 +
<tr>
 +
<th>Components</th>
 +
<th>Volume (ul)
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</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>EcoR1</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Pst1</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>0.5</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
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<td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Total</td>
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<td>10</td>
 +
</tr>
 +
</table>
 +
</p>
 +
 +
<p class="body_text">
 +
<b>INSERT IMAGE OF GEL 16/09 ZEO BB CANDIDATES DIGESTED WITH E&P FROM MINIPREPS OF COLONIES TRANSFORMED ON 13/09</b>
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</p>
 +
<p class="body_text">
 +
Prep digest of miniprep of J632014 with E and P in order to keep up the pSB1C3 stocks
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</p>
 +
 +
<p class="body_text">
 +
<table>
 +
<tr>
 +
<th>Components</th>
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<th>Cut</th>
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<th>Uncut</th>
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<th>Control - pSecTag2A</th>
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</tr>
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<tr>
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<td>J632014 DNA</td>
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<td>25</td>
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<td>5</td>
 +
<td>10</td>
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</tr>
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<tr>
 +
<td>EocR1</td>
 +
<td>2</td>
 +
<td>0</td>
 +
<td>0</td>
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</tr>
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<tr>
 +
<td>Pst1</td>
 +
<td>2</td>
 +
<td>0</td>
 +
<td>0</td>
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</tr>
 +
<tr>
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<td>Buffer 3</td>
 +
<td>4</td>
 +
<td>0</td>
 +
<td>0</td>
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</tr>
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<tr>
 +
<td>BSA</td>
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<td>0.5</td>
 +
<td>0</td>
 +
<td>0</td>
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</tr>
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<tr>
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<td>dH2O</td>
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<td>6.5</td>
 +
<td>5</td>
 +
<td>0</td>
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</tr>
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<tr>
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<td>Total</td>
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<td>40</td>
 +
<td>10</td>
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<td>10</td>
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</tr>
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</table>
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</p>
 +
 +
<p class="body_text">
 +
These were incubated for 2 hours at 37O C.
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Casey ran a gel of the following samples in the next order:
 +
Zeo BB: 26D, 26u; 28D, 28u; AuxD, AuxU <b>INSERT IMAGE OF GEL HERE PLS</B>
 +
</p>
 +
<p class="body_text">
 +
Gel1: E+P double digest and P, single digest (Auxin bb): 3, 12, 13, 15.
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Gel2: D digest 26, 28 and Auxin bb.
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Minipreps: 26, 28, 3, 12, 13, 15, 21 (sample of which DNA is waiting to be eluted)
 +
</p>
 +
 +
<p class="body_text">
 +
<b>Tuesday 17th September</b>
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</p>
 +
<p class="body_text">
 +
Miniprep of the 16 tubes of inoculations of MMP9 as well as tube 21 of which DNA had to be eluted.
 +
</p>
 +
<p class="body_text">
 +
Nanodrop results of the minipreps
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</p>
 +
 +
<p class="body_text">
 +
<table>
 +
<tr>
 +
<th>Tube no.</th>
 +
<th>ng/ul</th>
 +
<th>260/280</th>
 +
</tr>
 +
<tr>
 +
<td>1</td>
 +
<td>20.6</td>
 +
<td>1.9</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
 +
<td>26.9</td>
 +
<td>1.64</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
 +
<td>22.0</td>
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<td>3.2</td>
 +
</tr>
 +
<tr>
 +
<td>5</td>
 +
<td>4.8</td>
 +
<td>2.16</td>
 +
</tr>
 +
<tr>
 +
<td>6</td>
 +
<td>22.7</td>
 +
<td>2.99</td>
 +
</tr>
 +
<tr>
 +
<td>7</td>
 +
<td>16.4</td>
 +
<td>2.41</td>
 +
</tr>
 +
<tr>
 +
<td>8</td>
 +
<td>16.9</td>
 +
<td>2.01</td>
 +
</tr>
 +
<tr>
 +
<td>9</td>
 +
<td>7.1</td>
 +
<td>1.49</td>
 +
</tr>
 +
<tr>
 +
<td>10</td>
 +
<td>27.3</td>
 +
<td>1.79</td>
 +
</tr>
 +
<tr>
 +
<td>11</td>
 +
<td>16.0</td>
 +
<td>1.72</td>
 +
</tr>
 +
<tr>
 +
<td>12</td>
 +
<td>22.2</td>
 +
<td>1.41</td>
 +
</tr>
 +
<tr>
 +
<td>13</td>
 +
<td>11.0</td>
 +
<td>1.73</td>
 +
</tr>
 +
<tr>
 +
<td>14</td>
 +
<td>12.0</td>
 +
<td>1.6</td>
 +
</tr>
 +
<tr>
 +
<td>15</td>
 +
<td>18.4</td>
 +
<td>1.57</td>
 +
</tr>
 +
<tr>
 +
<td>17</td>
 +
<td>45.1</td>
 +
<td>1.62</td>
 +
</tr>
 +
<tr>
 +
<td>18</td>
 +
<td>9.2</td>
 +
<td>1.72</td>
 +
</tr>
 +
<tr>
 +
<td>21 (kc)</td>
 +
<td>21.3</td>
 +
<td>1.81</td>
 +
</tr>
 +
</table>
 +
</p>
 +
</div>
</div>

Revision as of 08:45, 4 October 2013

Lab Weeks

Week 16

Bacterial Labs

Results of the inoculations of transformations of 13/09 in 4xcmp: all showed growth apart from falcons 5.3, 5.2, 1.14 and 1.2. Glycerol stocks of the rest of 19 inoculations were made.

Minipreps of the above inoculations were made only for 1.3, 1.7, 1.9, 1.10, 1.11, 1.15, 1.20 and 5.1 due to lack of chromatographic columns.

Nanodrop result of the above minipreps showed concentration values below 16.0 ng/ul. Analytical digest of miniprep samples 1-9 (prepared the day before) as well as of 5.1 and 1.15 (which showed concentrations of about 15 ng/ul) with E and P

Components Volume (ul)
DNA 5
EcoR1 1
Pst1 1
Buffer 3 1
BSA 0.5
dH2O 1.5
Total 10

INSERT IMAGE OF GEL 16/09 ZEO BB CANDIDATES DIGESTED WITH E&P FROM MINIPREPS OF COLONIES TRANSFORMED ON 13/09

Prep digest of miniprep of J632014 with E and P in order to keep up the pSB1C3 stocks

Components Cut Uncut Control - pSecTag2A
J632014 DNA 25 5 10
EocR1 2 0 0
Pst1 2 0 0
Buffer 3 4 0 0
BSA 0.5 0 0
dH2O 6.5 5 0
Total 40 10 10

These were incubated for 2 hours at 37O C. Casey ran a gel of the following samples in the next order: Zeo BB: 26D, 26u; 28D, 28u; AuxD, AuxU INSERT IMAGE OF GEL HERE PLS

Gel1: E+P double digest and P, single digest (Auxin bb): 3, 12, 13, 15. Gel2: D digest 26, 28 and Auxin bb. Minipreps: 26, 28, 3, 12, 13, 15, 21 (sample of which DNA is waiting to be eluted)

Tuesday 17th September

Miniprep of the 16 tubes of inoculations of MMP9 as well as tube 21 of which DNA had to be eluted.

Nanodrop results of the minipreps

Tube no. ng/ul 260/280
1 20.6 1.9
2 26.9 1.64
4 22.0 3.2
5 4.8 2.16
6 22.7 2.99
7 16.4 2.41
8 16.9 2.01
9 7.1 1.49
10 27.3 1.79
11 16.0 1.72
12 22.2 1.41
13 11.0 1.73
14 12.0 1.6
15 18.4 1.57
17 45.1 1.62
18 9.2 1.72
21 (kc) 21.3 1.81