Gene Cloning: Successful!

Today, amid the project crises going on for my various classes, I got some very good and very exciting news:  my engineered microbial systems project seems to be going well!

A little background on the project and team:

Our team consists of three people:  Neal Singer (MechE), Jea Young Park (E:Bio) and myself (MechE).  Our professor is Jean Huang, who is awesome and exceedingly good at juggling multiple projects and inspiring us to do cool things.  The goal of our team was to (in the span of a month), clone a gene from p. Atlantica into e. Coli.  It sounds like it should be simple and routine, but it is actually quite a process.  The good news is it seems to have worked!

How do we know it worked?  Check out these gels:

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The “top” of the gel is side with the wells closest to the edge.  From left to right, we ran the PCR products from samples of plasmid from transformants in wells 1-9, and the 10th one was a positive control of PCR product of genomic DNA.  The idea is that if the plasmid contains the gene, we would see a band near the positive control which definitely contains a copy of the gene.  And the result is that we do!  This means that when we ligated the plasmid and the newly cloned gene, some of the copies re-circularized without the gene, and some of the copies incorporated the new gene, and some of those copies of the gene ended up in bacteria that we have growing in the lab.


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