Difference between revisions of "Team:Dundee/CGCraigon"

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            <span class="box-title">8/6: Amplification of <i>LbpA</i> into pQE80-L</span>
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                  <p><b>Aim of experiment:</b> To amplify <i>LbpA</i> for subsequent cloninng into the pQE80-L vector.</p>
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                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR </a></p>
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                  <p><b>Results:</b><a data-toggle="modal" data-target="#lbpa-figure2-modal" class="journal-protocol"> Figure 2 </a></p>
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<p><b> Next Steps:</b> The PCR product will be digested and ligated into pQE80-L tomorrrow.</p>
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            <span class="box-title">20/6: PCR of the pIDT-<i>OBP2A</i> plasmid using primers designed for the removal of the signalling peptide. </span>
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                <p><b>Aim of experiment:</b> To set up a PCR using pIDT-<i>OBP2A</i> using priers that if succesful will remove the signalling peptide from the n terminus of <i>OBP2A</i>. </p>
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                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#PCR-modal" class="journal-protocol"> PCR
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                  <p><b>Results:</b> N/A
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                  <p><b>Next Steps:</b> The next step will be to gel purify and extract the <i>OBP2A</i> PCR product from the PCR mixture. Ths will most likely be done tomorrow. 
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            <span class="box-title">21/6: Gel extraction of <i>OBP2A</i> from the PCR product and a confirmatory gel image of the purified <i>OBP2A</i>.  </span>
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                  <p><b>Aim of experiment:</b> To extract <i>OBP2A</i> from the PCR product using gel extraction. Then to subsequently image the extracted solution to determine the presence of <i>OBP2A<i>. </p>
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                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#?-modal" class="journal-protocol"> Gel Extraction
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                  <p><b>Results:</b> N/A
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                  <p><b>Next Steps:</b> The gel image shows a band corresponding to the size of <i>OBP2A</i>, The next step will be to restrict this gel extracted <i>OBP2A</i>in preperation for ligation.                </div>
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<!-- WEEK 2 -->
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        <div class="labtitle">Week Beginning 8/6/2015</div>
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          <p class="journal-summary-heading">Summary</p>
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        <p class="journal-content">Sequencing confirmed that <i>LbpA</i> had been successfully cloned into pSB1C3 therefore this week cloning was attempted to put <i>LbpA</i> into the high expression vector pQE80-L.</p>
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            <span class="box-title">8/6: Amplification of <i>LbpA</i> into pQE80-L</span>
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                  <p><b>Aim of experiment:</b> To amplify <i>LbpA</i> for subsequent cloninng into the pQE80-L vector.</p>
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                  <p><b>Results:</b><a data-toggle="modal" data-target="#lbpa-figure2-modal" class="journal-protocol"> Figure 2 </a></p>
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<p><b> Next Steps:</b> The PCR product will be digested and ligated into pQE80-L tomorrrow.</p>
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Revision as of 16:55, 6 September 2015

CSI: DUNDEE

The Forensic Toolkit

Watch Our Introduction Video
Nasal Mucus
Week Beginning 15/6/2015

Summary

Produced a plasmid preperation of pIDT-OBP2A in order to use it as template in PCR.

17/6: Transformation of pIDT-OBP2A in MC1610 E.coli strain.

Aim of Experiment: To transform pIDT-OBP2A into MC1061 e.coli strain

Protocols Used: Transformation

Results: N/A

Next Steps: If the transformation is successful the next step will be to produce an overnight culture for plasmid preparation.

18/6: Produce an overnight culture of pIDT-OBP2A plasmid in MC1061.

Aim of experiment: To produce an overnight culture of pIDT-OBP2A from positive colonies of MC1061 E.coli strain from the transformation done on the 17/6.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Produce a plasmid preparation of pIDT-OBP2A from this overnight culture.

19/6: Produce a plasmid preparation of pIDT-OBP2A from the overnight culture.

Aim of experiment: The overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain the pIDT-OBP2A plasmid.

Protocols Used: Miniprep

Results: The plasmid preperation yeilded a concentration of 305.53 ng/ul.

Next Steps: To run a PCR using the purified pIDT-OBP2A plasmid as a template for amplification.

Week Beginning 22/6/2015

Summary

Removal of the signaling peptide from the start of OBP2A and attempt to insert OBP2A into the biobrick vector pSB1C3.

17/6: Transformation of pIDT-OBP2A in MC1610 E.coli strain.

Aim of Experiment: To transform pIDT-OBP2A into MC1061 e.coli strain

Protocols Used: Transformation

Results: N/A

Next Steps: If the transformation is successful the next step will be to produce an overnight culture for plasmid preparation.

18/6: Produce an overnight culture of pIDT-OBP2A plasmid in MC1061.

Aim of experiment: To produce an overnight culture of pIDT-OBP2A from positive colonies of MC1061 E.coli strain from the transformation done on the 17/6.

Protocols Used: Overnight Cultures

Results: N/A

Next Steps: Produce a plasmid preparation of pIDT-OBP2A from this overnight culture.

19/6: Produce a plasmid preparation of pIDT-OBP2A from the overnight culture.

Aim of experiment: The overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain the pIDT-OBP2A plasmid.

Protocols Used: Miniprep

Results: The plasmid preperation yeilded a concentration of 305.53 ng/ul.

Next Steps: To run a PCR using the purified pIDT-OBP2A plasmid as a template for amplification.