Week 9, Semester 2.

Colorized low-temperature electron micrograph, E. coli. https://www.flickr.com/photos/microbeworld/5981923914/

Welcome back to this blog!

This week's focus consists of running a polymerase-chain reaction (PCR) on the DNA samples previously discussed in the Week 5, Semester 2 edition of this blog. It remains to be seen if the primers utilized were successful at the DNA amplification process, for as of publication time the PCR was still in progress; thus the results had not been analyzed.

The next phase of this project will proceed beginning next week with the extension of the extraction protocol to other bacterial species endemic to this lab. The methodology utilized for E. coli extractions and testing will be repeated with each additional species; every phase of the project from extraction, to electrophoresis gel, to spectrometer reading, to finally PCR will be repeated with each additional species to identify any correlations or errors in the methodology used and results obtained. 

After the multi-species extractions have been finished, the project will conclude for the semester with an analysis of the genomic sequences specific to each bacterial species. The primers selected for the initial E. coli PCR's were referenced as being universal in the work Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes (Frank, et. al, 2008) and were therefore selected for further testing. However, it is hypothesized that further examination of the individual genomes is warranted to determine if a new primer design is needed for application to species other than E. coli.

Further details to follow on the next blog issue.