Genome sequencing is a technique that identifies the order of nucleotides in a DNA sequence of an organism. This process is the most successful for large animals like humans and plants, however, genome sequencing of microbial eukaryotes is still advancing. Microbial Eukaryotes like Amoeba have very diverse and complex genomes making it more difficult to sequence than other larger organisms. The goal of this research is to collect pure DNA from amoebozoans for genome sequencing. The challenge is that amoeba live with bacteria and other small eukaryotes that make it difficult to discern amoeba DNA from others before sequencing. This relative contamination is problematic because these other organisms usually have simple genomes that are easily sequenced in comparison to amoeba with complex genomes. This complicates the process and cost of genome sequencing in amoebae. In this study, two methods are considered to isolate the cells for pure DNA extraction including antibiotic treatment and centrifugation including density gradient centrifugation and nuclear isolation. Preliminary results of the antibiotic treatment proved to be ineffective because the concentrations of the antibiotic used more detrimental to the cells of amoebae than the bacteria. DNA isolation based on centrifugation showed promising result. Development of a protocol for pure genomic DNA isolation will be useful in other research fields that depend on pure genomic DNA.
