Week 9- Accomplished Sharp Goals

Last week I accomplished my initial goal at Sharp.  I set out to show the applicability of these impedance sensors to be used on genomic assays.  To do this, I used three E. coli genes: adenylate kinase (ADK), hemolysin A (HlyA) and shiga toxin 2 (Stx2).  The sensors I ran my experiments on were functionalized with ADK DNA probes and I measured good data by a high specific ADK signal, and a low non-specific Hly and STX signal.  After repeating various experiments, and optimizing four parameters at which to run the sensors at, I successfully showed that these impedance sensors can be used to detect specific pathogenic bacterial species.

Now, I am running experiments on sensors functionlized with either Hly or STX to make sure that these sensors can be applied in variable ways. 

My second goal at Sharp is to compare single-stranded DNA and double-stranded DNA targets on the sensors.  ssDNA targets are primarily used, but if I can show that dsDNA targets produce similar results to single-stranded targets, then it will be more economically viable and will reduce steps (labeling, digesting with enzyme, and re-purifying).  However, this week I ran PCR twice in order to make single-stranded targets and the gel electrophoresis showed that PCR did not work either time.  I therefore have been stunted in my attempt to create single-stranded targets to compare alongside of the double-stranded targets I have already created.

-Vena