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>strain </span> | + | <span class="box-title">17/6: Transformation of pIDT-<i>OBP2A</i> in MC1610 <i>E.coli </i>strain. </span> |
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| + | <span class="box-title">17/6: Transformation of pIDT-<i>OBP2A</i> in MC1610 <i>E.coli </i>strain. </span> | ||
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| + | <p><b>Aim of Experiment:</b> To transform pIDT-<i>OBP2A</i> into MC1061 <i> e.coli</i> strain </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>Next Steps:</b> If the transformation is successful the next step will be to produce an overnight culture for plasmid preparation. </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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| + | <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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| + | <span class="box-title">19/6: Produce a plasmid preparation of pIDT-<i>OBP2A</i> from the overnight culture. </span> | ||
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| + | <p><b>Aim of experiment:</b> The overnight culture produced yesterday will now undergo a plasmid preparation in order to obtain the pIDT-<I>OBP2A</I> plasmid.</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>Next Steps:</b> To run a PCR using the purified pIDT-<i>OBP2A</i> plasmid as a template for amplification. | ||
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Revision as of 16:53, 6 September 2015
Watch Our Introduction Video
Summary
Produced a plasmid preperation of pIDT-OBP2A in order to use it as template in PCR.
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.
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.
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.
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 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.
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.
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.
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.
