EMCC Abstract

1. Why research Staphylococcus aureus DNA extraction?

In microbiology, experimental research is important because it allows us to reinforce what we already know about organisms, but it can also open the door to new discoveries. For example, defining the best technique to extract DNA material from an organism can either confirm a hypothesis made on existing knowledge or it can lead to the discovery of new information. Specifically, determining the best method to extract DNA from microorganisms, such as bacteria, can be found valuable to an academic institution, as it can serve as a basis for the development of further investigations made on that subject. For this experiment, protocols will be conducted to extract DNA material from the Staphylococcus aureus bacteria cells. Staphylococcus aureus or S. aureus for short is a bacterium that can be part of the normal skin flora of human beings and never cause a problem; the CDC calculates that 33% of Americans carry this microbe without illness (MRSA, 2016). However, if given the opportunity S. aureus will attack the body and cause illness; and what’s worse, is that many strains have developed resistance to widely used antibiotics such as Vancomycin and Methicillin making this seemingly harmless organism deadly under certain conditions (Gram-positive, 2011). While DNA extraction and analysis can be utilized for bacterial and strain identification it can also serve to help develop a solution for the surging worldwide problem of drug-resistant microorganisms, such as S. aureus, tallying up another important reason to research micro-DNA extraction.

2. Research tactics.

There will be a total of six DNA extraction protocols, which are: the Boiling Protocol, TE Boiling Protocol, TE Freeze and Thaw Protocol, Lysis Protocol, and Meat Tenderizer Protocol  (Haberkorn, James, & Leonetti, 2016), measuring up to a commercial DNA extraction kit guaranteed to render high yields of optimal quality DNA material (Ready-LyseTM, 2012). The extracted nuclei acid will be analyzed using a microvolume UV-Vis spectrophotometer to qualitatively quantify results. The values of interest that will be sought are the concentration of nucleic acid measured by the absorbance of light and also the quality of the material extracted calculated by the 260/280 ratio which is the nucleic acid ratio of absorbance at the wavelengths of 260nm and 280nm respectively (260/280, 2008).

4. Overview of findings

Research question- When comparing the quantity and quality of the DNA material extracted with the previously mentioned protocols, which DNA extraction protocol worked better in comparison to the commercial DNA extraction kit? Which one extracted higher yields? Which one extracted better quality DNA?

Hypothesis- My hypothesis is that the Meat Tenderizer Protocol will extract better quality DNA in higher yields, because of the intricacy of the steps involved in the process.

References

Gram-positive bacteria. (2011). In K. Rogers (Ed.), Bacteria and viruses. Retrieved from Gale Virtual Reference Library database. (Accession No. GALE|CX4040900042)

Haberkorn, M., James, J., & Leonetti, C. (20016). Developing an effective e-coli protocol. Raw data in preparation, Phoenix College, Phoenix, AZ.

MRSA tracking. (2016, march 3). In Centers for disease control and prevention. Retrieved December 16, 2016, from https://www.cdc.gov/mrsa/tracking/

Ready-LyseTM lysozyme solution [Pamphlet]. (2012). Retrieved from http://www.epibio.com/docs/default-source/protocols/ready-lyse-lysozyme-solution.pdf?sfvrsn=6

260/280 and 260/230 ratios [Pamphlet]. (2008). Retrieved from http://www.nanodrop.com/Library/T009-NanoDrop%201000-&-NanoDrop%208000-Nucleic-Acid-Purity-Ratios.pdf