Genome Assembly

One-Sentence Definition

Genome assembly reconstructs contiguous DNA sequences (contigs/scaffolds/complete chromosomes) from overlapping sequencing reads.

Simple Explanation

Reads are puzzle pieces; assembly puts the microbial genome picture together.

Detailed Scientific Explanation

  • De novo (SPAdes, Unicycler) vs reference-guided mapping
  • Short reads: high accuracy bases, fragmented plasmids/repeats
  • Long reads (ONT/PacBio): better continuity, need polishing
  • Hybrid assembly: best of both for many bacterial genomes
  • Metrics: N50, completeness (CheckM), contamination

Mechanism

Graph of overlaps (de Bruijn / overlap graphs) → path through graph → contigs; unresolved repeats create breaks — common at rRNA and insertion sequences.

Clinical Importance

  • Incomplete plasmid assembly → missed carbapenemase context
  • Mis-assembly can create false gene fusions (rare but important QC)

Research Importance

  • Closed genomes for reference; mobile element architecture

Diagnostic Relevance

AMR Relevance

Active Recall Questions

  1. Short-read vs long-read tradeoff?
  2. What genomic features break assemblies?
  3. Why does plasmid closure matter for CRE epi?

Connections