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Team:St Andrews/Procedure
From 2012.igem.org
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<dd><p><ul>Δ15 and Δ6 desaturases from <i>Synechocystis</i> in pET-15b were sent off for sequencing. Once again, the results were discouraging even though protein and lipid analysis showed the opposite. BLAST showed that the gene found in both samples corresponded to a <i>T. brucei</i> HMG-CoA reductase which we thought we had cut out from the plasmid. We thus performed a PCR, using the vectors sent off for sequencing as a template.</ul></p> | <dd><p><ul>Δ15 and Δ6 desaturases from <i>Synechocystis</i> in pET-15b were sent off for sequencing. Once again, the results were discouraging even though protein and lipid analysis showed the opposite. BLAST showed that the gene found in both samples corresponded to a <i>T. brucei</i> HMG-CoA reductase which we thought we had cut out from the plasmid. We thus performed a PCR, using the vectors sent off for sequencing as a template.</ul></p> | ||
| - | <p><ul><a data-toggle="modal" href="#modal7" class="btn btn-primary btn-mini">Sequence</a> | + | <p><ul><a data-toggle="modal" href="#modal7" class="btn btn-primary btn-mini"><font color="white">Sequence</font></a> |
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<dt>16/7/12 Δ6</dt> | <dt>16/7/12 Δ6</dt> | ||
| - | <dd><p><ul><a data-toggle="modal" href="#modal8" class="btn btn-primary btn-mini">Sequence</a> | + | <dd><p><ul><a data-toggle="modal" href="#modal8" class="btn btn-primary btn-mini"><font color="white">Sequence</font></a> |
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<dd><p><ul>After showing successful insertion of <i>L. major</i> Δ6 elongase in pET-15b, the sample was sent off for sequencing. Unfortunately, the gene sequenced was not the Δ6 elongase we were hoping to find, but a <i>T. brucei</i> mevalonate kinase, which was previously inserted in the pET-15b vectors we had been using. Thus, we digested some fresh pET-15b to be used in the future.</ul></p> | <dd><p><ul>After showing successful insertion of <i>L. major</i> Δ6 elongase in pET-15b, the sample was sent off for sequencing. Unfortunately, the gene sequenced was not the Δ6 elongase we were hoping to find, but a <i>T. brucei</i> mevalonate kinase, which was previously inserted in the pET-15b vectors we had been using. Thus, we digested some fresh pET-15b to be used in the future.</ul></p> | ||
| - | <p><ul><a data-toggle="modal" href="#modal6" class="btn btn-primary btn-mini">Sequence</a> | + | <p><ul><a data-toggle="modal" href="#modal6" class="btn btn-primary btn-mini"><font color="white">Sequence</font></a> |
<div class="modal hide" id="modal6"> | <div class="modal hide" id="modal6"> | ||
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Revision as of 12:07, 2 August 2012
Lab Book
Procedure
- date
- what we did
- date
- what we did
- date
- what we did
anyone really keen to write this up, up to protein visualization? Don't forget to mention the differing restriction enzymes and vectors of each team. I can add in links to the protocols page.
Please also refer to our Protocols page.
Sequences Primers Sequencing results Lipid analysis
Start from lipid extraction
primers, sequence results
Sequences
who knows what those Metal Mickies are up to. Eating nachos in the Whey Pat most likely. MEGA LOLS (all caps, with a Z).
Primers Digestion Transformation
Primers
All primers are notated 5' to 3'.
Please refer to the Primer annealing lab book entry for further detail.
