PacBio-Based Protocol for Bacterial Genome Assembly

Methods Mol Biol. 2021:2242:3-14. doi: 10.1007/978-1-0716-1099-2_1.

Abstract

Acquisition of high-quality bacterial genomes is fundamental, while having in mind investigation of subtitle intraspecies variation in addition to development of sensitive species-specific tools for detection and identification of the pathogens. In this view, Pacific Biosciences technology seems highly tempting taking into consideration over 10,000 bp length of the generated reads. In this work, we describe a bacterial genome assembly pipeline based on open-source software that might be handled also by non-bioinformaticians interested in transformation of sequencing data into reliable biological information. With the use of this method, we successfully closed six Dickeya solani genomes, while the assembly process was run just on a slightly improved desktop computer.

Keywords: Dickeya spp.; Next generation sequencing; Pectobacterium spp.; Single-molecule real-time sequencing; Soft rot bacteria; Whole-genome sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / genetics*
  • Databases, Genetic
  • Dickeya / genetics*
  • Genome, Bacterial*
  • Genomics*
  • High-Throughput Nucleotide Sequencing
  • Research Design
  • Single Molecule Imaging
  • Whole Genome Sequencing
  • Workflow

Substances

  • DNA, Bacterial