Difference between revisions of "Team:Dundee/CGCraigon"
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| − | <span class="box-title">17/6: Transformation of pIDT-<i>OBP2A</i> in MC1610 <i>e.coli </i> | + | <span class="box-title">17/6: Transformation of pIDT-<i>OBP2A</i> in MC1610 <i>e.coli </i>strain </span> |
<span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-1"></span> | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-1"></span> | ||
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<p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformation </a> </p> | <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#transformation-modal" class="journal-protocol">Transformation </a> </p> | ||
<p><b>Results: </b><a data-toggle="modal" data-target="#lbpa-figure1-modal" class="journal-protocol"> N/A </a></p> | <p><b>Results: </b><a data-toggle="modal" data-target="#lbpa-figure1-modal" class="journal-protocol"> N/A </a></p> | ||
| − | <p><b>Next Steps:</b> If the transformation is successful the next step will be to produce overnights in preperation for | + | <p><b>Next Steps:</b> If the transformation is successful the next step will be to produce overnights in preperation for plasmid perpetration. </p> |
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| − | <span class="box-title">19/6: Plasmid | + | <span class="box-title">19/6: Plasmid preparation of pIDT-<i>OBP2A</i> overnight culture from yesterday. </span> |
<span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-3"></span> | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-3"></span> | ||
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| − | <p><b>Aim of experiment:</b> | + | <p><b>Aim of experiment:</b> the overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain pIDT-<I>OBP2A</I> that can then be used as template in PCR. </p> |
<p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Miniprep </a></p> | <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#miniprep-modal" class="journal-protocol"> Miniprep </a></p> | ||
<p><b>Results:</b> The plasmid preperation yeilded a concentration of 305.53 ng/ul. | <p><b>Results:</b> The plasmid preperation yeilded a concentration of 305.53 ng/ul. | ||
| − | <p><b>Next Steps:</b> A PCR using the purified pIDT-<i>OBP2A</i> | + | <p><b>Next Steps:</b> A PCR using the purified pIDT-<i>OBP2A</i> as a plasmid template can now be done. |
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Revision as of 16:19, 6 September 2015
Watch Our Introduction Video
Summary
Produce a plasmid preperation of OBP2A in the IDT plasmid
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 overnights in preperation for plasmid perpetration.
Aim of experiment: To produce overnights of pIDT-OBP2A in MC1061.
Protocols Used: Overnight Cultures
Results: N/A
Next Steps: A plasmid preperation of pIDT-OBP2A will be done next.
Aim of experiment: the overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain pIDT-OBP2A that can then be used as template in PCR.
Protocols Used: Miniprep
Results: The plasmid preperation yeilded a concentration of 305.53 ng/ul.
Next Steps: A PCR using the purified pIDT-OBP2A as a plasmid template can now be done.
Summary
Removal of the signaling peptide from the start of OBP2A and attempt to insert OBP2A into the biobrick vector pSB1C3.
Aim of experiment: To set up a PCR using pIDT-OBP2A using priers that if succesful will remove the signalling peptide from the n terminus of OBP2A.
Protocols Used: PCR
Results: N/A
Next Steps: The next step will be to gel purify and extract the OBP2A PCR product from the PCR mixture. Ths will most likely be done tomorrow.
Aim of experiment: To extract OBP2A from the PCR product using gel extraction. Then to subsequently image the extracted solution to determine the presence of OBP2A.
Protocols Used: Gel Extraction
Results: N/A
Next Steps: The gel image shows a band corresponding to the size of OBP2A, The next step will be to restrict this gel extracted OBP2Ain preperation for ligation.
Summary
Sequencing confirmed that LbpA had been successfully cloned into pSB1C3 therefore this week cloning was attempted to put LbpA into the high expression vector pQE80-L.
