Difference between revisions of "Team:Dundee/CGCraigon"

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             <span class="box-title">23/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061.  </span>
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             <span class="box-title">23/6: Gel extraction of <I>OBP2A</I> from the PCR performed on the 22/6.  </span>
 
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                  <p><b>Aim of experiment:</b> To perform a gel extraction of OBP2A from a PCR reaction mixture that was performed on the 22/6. Once gel extraction has been performed a subsequent gel will be done to determine if the gel extraction has been successful. 
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                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#gel extraction -modal" class="journal-protocol">Gel extraction </a></p>
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        <p><b>Results:</b> as can be seen from the gel image a band appears at 450b base pairs corresponding to the size of the <I>OBP2A</I> gene fragment.
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                  <p><b>Next Steps:</b> To perform a restriction of <I>OBP2A</I> in preparation for ligation.              </div>
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            <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061.  </span>
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                   <p><b>Aim of experiment:</b> To produce an overnight culture of pIDT-<i>OBP2A</i> from positive colonies of MC1061 <I>E.coli</I> strain from the transformation done on the 17/6.   
 
                   <p><b>Aim of experiment:</b> To produce an overnight culture of pIDT-<i>OBP2A</i> from positive colonies of MC1061 <I>E.coli</I> strain from the transformation done on the 17/6.   
 
                   <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
 
                   <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
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            <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061.  </span>
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              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-2"></span>
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                <div id="nmcollapsiblew1-2" class="collapse box-content">
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                  <p><b>Aim of experiment:</b> To produce an overnight culture of pIDT-<i>OBP2A</i> from positive colonies of MC1061 <I>E.coli</I> strain from the transformation done on the 17/6. 
 +
                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
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        <p><b>Results:</b> N/A
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                  <p><b>Next Steps:</b>  Produce a plasmid preparation of pIDT-<i>OBP2A</i> from this overnight culture.                </div>
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            <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061.  </span>
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              <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-2"></span>
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                <div id="nmcollapsiblew1-2" class="collapse box-content">
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                  <p><b>Aim of experiment:</b> To produce an overnight culture of pIDT-<i>OBP2A</i> from positive colonies of MC1061 <I>E.coli</I> strain from the transformation done on the 17/6. 
 +
                  <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Overnight Cultures </a></p>
 +
        <p><b>Results:</b> N/A
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                  <p><b>Next Steps:</b>  Produce a plasmid preparation of pIDT-<i>OBP2A</i> from this overnight culture.                </div>
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Revision as of 22:11, 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 OBP2A and insertion of OBP2A into the biobrick vector pSB1C3.

22/6: PCR of pIDT-OBP2A using primers designed for the removal of the signaling peptide.

Aim of Experiment: To perform a PCR on the plasmid preparation of pIDT-OBP2A produced last week, using primers that have been designed in a way as to remove the signalling peptide at the start of the OBP2A gene.

Protocols Used: PCR

Results: N/A

Next Steps: The next step will be to run the PCR product on a gel and perform a gel extraction on the part of the gel corresponding to the size of the OBP2A gene fragment.

23/6: Gel extraction of OBP2A from the PCR performed on the 22/6.

Aim of experiment: To perform a gel extraction of OBP2A from a PCR reaction mixture that was performed on the 22/6. Once gel extraction has been performed a subsequent gel will be done to determine if the gel extraction has been successful.

Protocols Used: Gel extraction

Results: as can be seen from the gel image a band appears at 450b base pairs corresponding to the size of the OBP2A gene fragment.

Next Steps: To perform a restriction of OBP2A in preparation for ligation.

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.

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.

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.