Difference between revisions of "Team:Dundee/lucylabjournal"
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| − | <p class="journal-content">faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. | + | <p class="journal-content">faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. .</p> |
</div> | </div> | ||
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<div class="border-day"> | <div class="border-day"> | ||
<div class="box"> | <div class="box"> | ||
| − | <span class="box-title"> | + | <span class="box-title">8/6: Amplification of LbpA for Cloning into pQE80-L</span> |
<span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#collapsiblew2-1"></span> | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#collapsiblew2-1"></span> | ||
<div id="collapsiblew2-1" class="collapse box-content"> | <div id="collapsiblew2-1" class="collapse box-content"> | ||
| − | <p><b>Aim of experiment:</b> | + | <p><b>Aim of experiment:</b> To amplify LbpA for subsequent cloninng into the pQE80-L vector.</p> |
| − | <p>Protocols Used: | + | <p>Protocols Used: PCR Protocol |
<p class="journal-content">faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.</p> | <p class="journal-content">faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.</p> | ||
</div> | </div> | ||
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<div class="modal-body"> | <div class="modal-body"> | ||
<p> | <p> | ||
| − | + | Restriction digests were set up according to the table below. The appropriate reaction and restriction enzymes that were used is stated in the <strong>Journal</strong> section. | |
</p> | </p> | ||
| − | + | <table border="1" cellspacing="0" cellpadding="0"> | |
| − | + | ||
| − | + | ||
| − | <table border="1" cellspacing="0" cellpadding="0 | + | |
<tbody> | <tbody> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | + | </td> | |
| − | <strong> | + | <td width="170" valign="top"> |
| + | <p> | ||
| + | <strong>Post Plasmid Purification</strong> | ||
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="151" valign="top"> |
| − | <p | + | <p> |
| − | <strong> | + | <strong>Post PCR</strong> |
| + | </p> | ||
| + | </td> | ||
| + | <td width="139" valign="top"> | ||
| + | <p> | ||
| + | <strong>gBlock</strong> | ||
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | DNA | + | <strong>DNA</strong> |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | 1mg | |
</p> | </p> | ||
</td> | </td> | ||
| − | + | <td width="151" valign="top"> | |
| − | + | <p> | |
| − | <td width=" | + | 50µl |
| − | <p | + | |
| − | + | ||
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="139" valign="top"> |
| − | <p | + | <p> |
| − | + | 100ng | |
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | + | <strong>Cutsmart Buffer (µl)</strong> | |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | 6 | |
</p> | </p> | ||
</td> | </td> | ||
| − | + | <td width="151" valign="top"> | |
| − | + | <p> | |
| − | <td width=" | + | 6 |
| − | <p | + | |
| − | + | ||
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="139" valign="top"> |
| − | <p | + | <p> |
| − | + | 6 | |
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | + | <strong>Water (µl)</strong> | |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | As required | |
| + | </p> | ||
| + | </td> | ||
| + | <td width="151" valign="top"> | ||
| + | <p> | ||
| + | 1 | ||
| + | </p> | ||
| + | </td> | ||
| + | <td width="139" valign="top"> | ||
| + | <p> | ||
| + | As required | ||
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | + | <strong>Enzyme 1 (µl)</strong> | |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | 1.5 | |
| + | </p> | ||
| + | </td> | ||
| + | <td width="151" valign="top"> | ||
| + | <p> | ||
| + | 1.5 | ||
| + | </p> | ||
| + | </td> | ||
| + | <td width="139" valign="top"> | ||
| + | <p> | ||
| + | 1.5 | ||
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | + | <strong>Enzyme 2 (µl)</strong> | |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | 1.5 | |
| + | </p> | ||
| + | </td> | ||
| + | <td width="151" valign="top"> | ||
| + | <p> | ||
| + | 1.5 | ||
| + | </p> | ||
| + | </td> | ||
| + | <td width="139" valign="top"> | ||
| + | <p> | ||
| + | 1.5 | ||
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td width=" | + | <td width="164" valign="top"> |
| − | <p | + | <p> |
| − | + | <strong><u>Total Volume (µl)</u></strong> | |
</p> | </p> | ||
</td> | </td> | ||
| − | <td width=" | + | <td width="170" valign="top"> |
| − | <p | + | <p> |
| − | + | 30 | |
| + | </p> | ||
| + | </td> | ||
| + | <td width="151" valign="top"> | ||
| + | <p> | ||
| + | 60 | ||
| + | </p> | ||
| + | </td> | ||
| + | <td width="139" valign="top"> | ||
| + | <p> | ||
| + | 30 | ||
</p> | </p> | ||
</td> | </td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
| − | </table> | + | </table> |
<p> | <p> | ||
| − | + | <img | |
| + | width="628" | ||
| + | height="34" | ||
| + | src="file:///C:/Users/Lucy/AppData/Local/Temp/msohtmlclip1/01/clip_image001.png" | ||
| + | alt="Text Box: Table x: The components of the reaction digest reactions and their respective quantities." | ||
| + | /> | ||
| + | <strong><u></u></strong> | ||
</p> | </p> | ||
<p> | <p> | ||
| − | + | <br/> | |
| + | The digests were incubated at 37<sup>o</sup>C for 3 hours. | ||
| + | <br/> | ||
| + | Note that when performing plasmid digestions, 2.5µl of alkaline phosphatase and 2.5µl of its buffer was added at the 2 hour and 2.5 hour mark. | ||
</p> | </p> | ||
| + | |||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 14:23, 15 July 2015
LABJOURNAL
BioSpray
Our forensic toolkit aims to use synthetic biology approaches to improve on the current methods used by crime scene investigators.
Summary
Lactoferrin binding protein A (LbpA) was successfully cloned into the vector pSB1C3.
Aim of Experiment: Amplify LbpA gene from N.meningitidis template.
Protocols Used: Click here to see our PCR protocol.
Results: Figure 1
Next Steps: Since the LbpA gene was amplified successfully, it was ready to be digested for subsequent cloning into pSB1C3.
Aim of experiment: To digest LbpA with BamHI and EcoRI and then ligate it into pSB1C3.
Protocols Used: Restriction Digests Ligations
Results: N/A
Next Steps: Ligations of LbpA + pSB1C3 were left overnight until the next day when they would be transformed into JM110 cells.
Aim of experiment: To transform the pSB1C3 + LbpA ligations into JM110 cells.
Protocols Used: Transformations
Results: N/A
Next Steps: Transformations will be checked tomorrow and if successful, overnight cultures will be set up.
Aim of experiment: To set up overnight cultures of the LbpA + pSB1C3 JM110 colonies.
Protocols Used: Overnight Cultures
Results: N/A
Next Steps: A miniprep will be performed on the overnight cultures and a pre-sequence digest test performed tomorrow.
Aim of experiment: To miniprep the LbpA + pSB1C3 JM110 overnight cultures and perform a pre-sequence restriction digest to check for the presence of the insert.
Protocols Used: Plasmid Purification (QIAprep® Spin Miniprep Kit) Restriction Digests
Results: N/A
Next Steps: Since the pre-sequencing digest worked, the miniprep will be sent for sequencing.
Week summary
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. .
Aim of experiment: To amplify LbpA for subsequent cloninng into the pQE80-L vector.
Protocols Used: PCR Protocol
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Restriction digests were set up according to the table below. The appropriate reaction and restriction enzymes that were used is stated in the Journal section.
|
Post Plasmid Purification |
Post PCR |
gBlock |
|
|
DNA |
1mg |
50µl |
100ng |
|
Cutsmart Buffer (µl) |
6 |
6 |
6 |
|
Water (µl) |
As required |
1 |
As required |
|
Enzyme 1 (µl) |
1.5 |
1.5 |
1.5 |
|
Enzyme 2 (µl) |
1.5 |
1.5 |
1.5 |
|
Total Volume (µl) |
30 |
60 |
30 |
The digests were incubated at 37oC for 3 hours.
Note that when performing plasmid digestions, 2.5µl of alkaline phosphatase and 2.5µl of its buffer was added at the 2 hour and 2.5 hour mark.
'