Rapid, user-friendly, cost-effective DNA and library Preparation methods for whole-genome sequencing of bacteria with varying cell wall composition and GC content using minimal DNA on the illumina platform.

ABSTRACT:

Background

Whole-genome sequencing using high-throughput sequencing is essential for identifying and characterising chromosomes and plasmids in nosocomial and environmental bacterial pathogens, including those with bioterrorism potential. To expedite outbreaks investigations, including accidental or intentional bacterial release, without compromising sequencing quality, we evaluated a more time-efficient, user-friendly, and cost-effective approach, using minimal DNA (~ 1 ng) from a single bacterial colony. Four DNA extraction methods were compared: the automated nucleic acid extractor (EZ1 Advanced, Qiagen) with or without DNA purification using AMPure^® beads (EZ1 vs. EZ1-AMP), and two rapid and inexpensive methods: heat shock lysis (HS), and glass bead disruption (GBD).

SUBMITTER: Bearzatto B 

PROVIDER: S-EPMC12020097 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature