"
Team:Cornell/testing/notebook/wetlab
From 2012.igem.org
(Difference between revisions)
| Line 41: | Line 41: | ||
<div class="row last-ele" style="margin-top:20px;"> | <div class="row last-ele" style="margin-top:20px;"> | ||
<div class="three columns offset-by-nine"> | <div class="three columns offset-by-nine"> | ||
| - | <a href="https://2012.igem.org/Team:Cornell/testing/notebook/wetlab/ | + | <a href="https://2012.igem.org/Team:Cornell/testing/notebook/wetlab/pre" style="display:block;float:right;">Bootcamp >></a> |
</div> | </div> | ||
</div> | </div> | ||
| Line 62: | Line 62: | ||
<div class="row last-ele"> | <div class="row last-ele"> | ||
<div class="three columns offset-by-nine"> | <div class="three columns offset-by-nine"> | ||
| - | <a href="https://2012.igem.org/Team:Cornell/testing/notebook/wetlab/ | + | <a href="https://2012.igem.org/Team:Cornell/testing/notebook/wetlab/pre" style="display:block;float:right;">Bootcamp >></a> |
</div> | </div> | ||
</div> | </div> | ||
| Line 79: | Line 79: | ||
$(this).hide(); | $(this).hide(); | ||
}) | }) | ||
| - | + | ||
}) | }) | ||
</script> | </script> | ||
</body> | </body> | ||
</html> | </html> | ||
Revision as of 20:23, 3 October 2012
Wet Lab Notebook Overview
-
Overview
We began the summer by holding a synthetic biology bootcamp in the DeLisa Lab. The purpose of this bootcamp was both to introduce new members to techniques in molecular biology and to get a running start on the cloning work for our project. During bootcamp, we successfully constructed both versions of our arsenic reporter, and attempted a Gibson assembly of a naphthalene-degrading plasmid.
In late June, we transitioned from bootcamp to our permanent bench space in Dr. Archer’s lab in Weill Hall. After spending a few weeks setting up the lab space troubleshooting general issues, we successfully constructed both versions of our salicylate reporter and began an alternative approach to construct a plasmid with a naphthalene-degrading (nah) operon. In parallel, we realized that electroporation efficiency for Shewanella transformation is less than optimal—to say the least. However, we were able to conjugate our constructs into Shewanella using a protocol provided by Dr. Gralnick.
As we transitioned into the fall semester in late August, wetlab work was divided into subprojects that could be accomplished in parallel. Subproject leaders independently worked on (1) characterizing our engineered Shewanella strains using reactors in the Angenent lab, (2) characterizing inducible promoters via qPCR of mtrB transcript in response to analyte, (3) characterizing inducible promoters in Shewanella via fluorescent reporters,(4) appending His-tags to MtrB in order to perform immunoassays, (5) performing site-directed mutagenesis on the nah operon to delete BioBrick cutsites, (6) constructing our final naphthalene-degrading plasmid to be conjugated into Shewanella, and (7) confirming that strains carrying the naphthalene-degrading plasmid can actually eat naphthalene.
