Purpose
To examine the microbial population of an elliptical at the Dixon Recreation Center by analysis of DNA from an uncultured community sample. 16S rRNA genes will distinguish bacteria and represent the microbial diversity of the sampled elliptical. An estimate of the cleanliness of the elliptical will be made based on species-abundance as well as evidence of any pathogenic strains. We will also compare and contrast respective populations of bacteria on clean and disinfected ellipticals to assess disinfectant effectiveness.
Methods
We collected bacteria from the elliptical, purified and amplified the total community DNA, and then analyzed the sequenced DNA with Mi Seq Software. A revised protocol was used to extract, purify, and sequence the community DNA; these methods were followed without deviation (1). The procedure to sequence and configure the data is described on Blackboard (2).
Results

PCR amplification of the community DNA product was unsuccessful. There was no evidence of PCR amplified DNA on the gel (Figure 1.1).

An alternate gel from Molecular Microbiology Laboratory was used as a model to perform gel electrophoresis of the community DNA (2). A plot of migration distance vs. size that includes the Invitrogen ladder can be used to determine the size of the PCR amplified community DNA (Figure 1.2).

[Figure 1.2] Base Pairs vs. Migration Distance of Invitrogen Low DNA Mass Ladder. The distance these standardized fragments is associated with a quantity of base pairs. The graph created from a plot of base pairs vs. migration distance produces an equation that can be used to determine the size of the PCR amplified community DNA.

The length of the amplicon was determined by using lane 36 of the alternate gel (2). The estimated migration distance of the amplicon in lane 36 is