De Novo Genome Sequencing, Annotation, and Taxonomy of Unknown Bacteria

Abstract

Whole-genome sequencing of viruses and bacteria has become routine thanks to advances in DNA-sequencing technologies. Parallel advances in computing power and software design allow for billions of base pairs of sequence information to be analyzed in hours to minutes. Here, I describe methods to isolate known as well as new species of bacteria from the environment; to purify, sequence, assemble, and bioinformatically annotate their genomes; and to determine their place in the tree of life by phylogenetic analysis. The protocol introduced here was developed as part of Cold Spring Harbor’s Advanced Bacterial Genetics course.