Research

What to know:

As a student seeking to go into Chemical Engineering, and someone who has an avid interest in microbiology/DNA, I began this internship with Aeon Global Health with a question: What factors affect DNA Extraction and Purification? Answering this question is very important to the Genetics industry and may create protocols that yield a much better DNA Extraction and Purification. Based on this answer, we may be able to streamline (or at least assure the current protocol is the best way) the protocol and yield even better results.

What I Know about the Topic:

For my testing, I used 4 variables so as to test: Room Temperature Storage, 50 Degrees Celsius Storage, Acidity (specifically 3.5 pH) and Salinity (although I did not measure the pH of the Salt Water) along with a Control Swab of my own DNA and Control Sample from the RNaseP DNA Kit. Given some preconceived notions, I predicted that the High Temperature Storage and the Acidity samples will yield the least amount of DNA from the DNA Extraction. For the Purification process, I predict that the Salinity, Acidity and High Temperature Storage Samples will have the lowest concentration of Purified DNA that is able to be analyzed.

The Process:

Beginning with the samples, I created a plate map (well assignments for the Swabs) and began a series of washes so as to pull the Saliva from the Swab. Next, I took the extracted Saliva and ran it through an Elution Buffer which allowed the DNA in the Saliva to precipitate through and allowed for me to extract it, bind magnetic beads to it and wash it so as to leave Purified DNA. From there, I was able to splice and replicate the DNA in the Polymerase Chain Reaction Machine and then put the Replicated DNA into a chip to be analyzed by a Computer Program. From there, I went in and analyzed the DNA, correcting Discordant "calls" and creating the DNA Profile for my Pharmacogenetic Report.

Results:

For the Extraction Step, I was correct. The Acidity and High Temperature Storage Samples yielded the least concentration of DNA. Surprisingly, however, the DNA Analysis showed the Salinity Sample had no discordant "calls" and had a 100% Confidence "Call" rate, at least 2-5% higher than both Control DNA Samples. As it would appear, while the Extraction DNA Concentration is a bit lower for the Salinity Sample, the Analysis results are far superior than the original Protocol.