DNA Extraction Methods on Enterococcus Faecalis (Week 3)

During my 3rd week, I began working on my research project, which involved 4 DNA extraction methods following a set of protocols. I am trying to determine which DNA extraction method will yield the most of amount of DNA from Enterococcus faecalis. The methods that I am testing include a commercial kit and 3 (one includes 3 different chemicals to use) methods that I chose. The name of the commercial kit is QuickExtract. For this method, I followed a protocol and used QuickExtract Bacterial DNA and Ready-Lyse Lysozyme (1st image shown below). Since it is a commercial kit, it is supposed to yield the most amount of DNA. As for the other 3 methods, I will use protocols for a boiling, microwave and enzymatic digestion followed by heating (done using 3 different chemicals) method. The boiling method did not require any chemicals so I only needed my bacteria, a centrifuge and a water bath. For the microwave method, I will need a centrifuge, TE buffer (2nd image shown below), 15% SDS, a microwave, lysis buffer and ethanol. As for the enzymatic digestion followed by heating method, a centrifuge is needed along with 3 different chemicals (only that each time I perform the protocol, only one chemical is used). The 3 different chemicals that I will use are Ready-Lyse Lysozyme, proteinase K, 10% Tween 20, and 15% SDS (proteinase K, 10% Tween 20, and 15% SDS will be used altogether in during one protocol). However, during this week, I was only able to perform 3 methods: QuickExtract, boiling and one of the ways to perform enzymatic digestion followed by heating (I used Ready-Lyse Lysozyme). Next week, I will be working on the microwave method and using the remaining chemicals for the enzymatic digestion followed by heating method.

QuickExtract Bacterial DNA & Ready-Lyse Lysozyme

TE buffer