Comparative Analysis of DNA Isolation Methods for the Human Microbiome

Undergraduate Honors Thesis

Citations

Citeable URL: https://scholar.colorado.edu/concern/undergraduate_honors_theses/zw12z592q

Abstract

The human microbiome is made up of millions of microbes coexisting in and on us with involvement in health and disease. The exact mechanisms by which this occurs have yet to be fully elucidated. Due to high variability in the composition of each person’s microbiome, it requires large numbers of people to begin to identify some of these processes. A major road block to generating large data sets for analysis is the high cost of DNA isolation from large collections of phenotypically well-characterized human subjects. This study explores the possible application of an inexpensive alternative DNA isolation method utilizing hot alkaline-ethanol lysis (known as EtNa), in the large diverse US cohort assembled in the Adolescent to Adult Health Study (ADD Health). Based on alpha and beta diversity analysis, EtNa is shown to be similar to more standard protocols but is clearly not identical. We report results of this comparison as well as some preliminary characterizations of the ADD Health microbiome that will ultimately be used in a larger study.

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