Difference between revisions of "Team:Sheffield/Notebook"
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<b> <u> Week 3: </b> </u> <br> | <b> <u> Week 3: </b> </u> <br> | ||
| − | - | + | -Ensured we had sufficient amounts of plasmid (pSB1C3, pUC18 and pBSKII) for further cloning experiments by transforming them into Top10 competent cells and re-harvesting them (mini-prep experiments) <br> |
| − | - | + | -Characterised the plasmid stocks obtained (Nano Drop and agarose gel)<br> |
| − | - | + | -Characterised the genotype of wild-type (W3110) and mutant (JC28) strains:<br> |
| − | + | -genomic DNA extractions have been performed <br> | |
| − | + | - <i>entC</i> gene has been amplified- PCR <br> | |
| − | + | -PCR products have been analysed- agarose gel </p> | |
<h3> Constitutive hemerythrin expression </h3> | <h3> Constitutive hemerythrin expression </h3> | ||
Revision as of 20:49, 18 October 2016
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NOTEBOOK |
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CLONING OF HEMERYTHRIN
Week 1:
-Literature research
-Decided an experimental outline
-In silico design of hemerythrin constructs
-Synthetic genes and primers required for cloning have been ordered
Week 2:
-Make competent cells for the E.coli strains that we used during our project- Top10, JC28 and W3110
-Tested the efficiency of transformation in these strains
-Prepared buffers, growth media and pouring agar plates for the experiments we were planning to carry out during the following weeks
-Several experiments have been performed in order to monitor the growth of the wild type (W3110) and siderophore-deficient mutant (JC28)
Week 3:
-Ensured we had sufficient amounts of plasmid (pSB1C3, pUC18 and pBSKII) for further cloning experiments by transforming them into Top10 competent cells and re-harvesting them (mini-prep experiments)
-Characterised the plasmid stocks obtained (Nano Drop and agarose gel)
-Characterised the genotype of wild-type (W3110) and mutant (JC28) strains:
-genomic DNA extractions have been performed
- entC gene has been amplified- PCR
-PCR products have been analysed- agarose gel
