Difference between revisions of "Team:Dundee/CGCraigon"
Line 126: | Line 126: | ||
<div class="border-day"> | <div class="border-day"> | ||
<div class="box"> | <div class="box"> | ||
− | <span class="box-title">23/6: | + | <span class="box-title">23/6: Gel extraction of <I>OBP2A</I> from the PCR performed on the 22/6. </span> |
<span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew2-2"></span> | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew2-2"></span> | ||
<div id="nmcollapsiblew2-2" class="collapse box-content"> | <div id="nmcollapsiblew2-2" class="collapse box-content"> | ||
+ | <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. | ||
+ | <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#gel extraction -modal" class="journal-protocol">Gel extraction </a></p> | ||
+ | <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. | ||
+ | |||
+ | |||
+ | <p><b>Next Steps:</b> To perform a restriction of <I>OBP2A</I> in preparation for ligation. </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="row"> | ||
+ | <div class="border-day"> | ||
+ | <div class="box"> | ||
+ | <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061. </span> | ||
+ | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-2"></span> | ||
+ | <div id="nmcollapsiblew1-2" class="collapse box-content"> | ||
<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> | ||
Line 138: | Line 153: | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | <div class="row"> | ||
+ | <div class="border-day"> | ||
+ | <div class="box"> | ||
+ | <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061. </span> | ||
+ | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-2"></span> | ||
+ | <div id="nmcollapsiblew1-2" class="collapse box-content"> | ||
+ | <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 | ||
+ | |||
+ | |||
+ | <p><b>Next Steps:</b> Produce a plasmid preparation of pIDT-<i>OBP2A</i> from this overnight culture. </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <div class="row"> | ||
+ | <div class="border-day"> | ||
+ | <div class="box"> | ||
+ | <span class="box-title">18/6: Produce an overnight culture of pIDT-<i>OBP2A</i> plasmid in MC1061. </span> | ||
+ | <span type="button" class="chevron_toggleable box-btn glyphicon glyphicon-chevron-down" data-toggle="collapse" data-target="#nmcollapsiblew1-2"></span> | ||
+ | <div id="nmcollapsiblew1-2" class="collapse box-content"> | ||
+ | <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 | ||
+ | |||
+ | |||
+ | <p><b>Next Steps:</b> Produce a plasmid preparation of pIDT-<i>OBP2A</i> from this overnight culture. </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
Revision as of 22:11, 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 OBP2A and insertion of OBP2A into the biobrick vector pSB1C3.
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.
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.
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: 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: 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.