Difference between revisions of "Team:Dundee/CGCraigon"
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| − | <p><b>Next Steps:</b>If the transformation is successful and positive colonies form on the agar plate then these colonies will | + | <p><b>Next Steps:</b>If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep. </div> |
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| − | <p><b>Aim of experiment:</b> | + | <p><b>Aim of experiment:</b> The transformation performed on the 24/6 failed and as a result <I>OBP2A</I> will again ligated into the biobrick vector pSB1C3 and then transformed into JM110 <I>E.coli</I> strain. |
| − | <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol"> | + | <p><b>Protocols Used:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Ligation:</b> <a data-toggle="modal" data-target="#onc-modal" class="journal-protocol">Transformation</a> </a></p> |
<p><b>Results:</b> N/A | <p><b>Results:</b> N/A | ||
| − | <p><b>Next Steps:</b> | + | <p><b>Next Steps:</b>If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep. </div> |
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Revision as of 23:17, 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 it can be seen from the gel image, a band is present 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 firstly restrict the gel extracted OBP2A. Secondly, to then ligate the restricted OBP2A into the biobrick vector pSB1C3. Third and finally to transform this plasmid into the JM110 E.coli strain.
Protocols Used: Restriction Digest : Ligation : Transformation
Results: N/A
Next Steps:If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep.
Aim of experiment: The transformation performed on the 24/6 failed and as a result OBP2A will again ligated into the biobrick vector pSB1C3 and then transformed into JM110 E.coli strain.
Protocols Used: Ligation: Transformation
Results: N/A
Next Steps:If the transformation is successful and positive colonies form on the agar plate then these colonies will then be grown overnight in preparation for Miniprep.
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.
