A practical introduction to bacterial genomics
5 important questions on A practical introduction to bacterial genomics
What are the steps to take to get to the whole-genome sequencing of bacteria?
- isolate bacteria from clinical specimens, healthy people or environment
DNA isolation
- Release DNA from bacterial cells
- Purify DNA to a high concentration
Sequence library preparation
- Prepare DNA for sequencing
- fragment DNA to get correct length
- ligate barcodes and adapters
Whole-genome sequencing
- Determine the base sequence of the DNA molecules
How does Illumina sequencing work?
- PCR-based method
- Short, paired-end reads with high accuracy
How does Nanopore sequencing work?
- No PCR needed
- long, single-end reads with many errors
- 90-95% accuracy
- Higher grades + faster learning
- Never study anything twice
- 100% sure, 100% understanding
What is "de novo assembly"?
- Try to stitch together all sequencing reads
- Create a genome from scratch/de novo using Unicycler software
What is the problem with repeats in the genome?
By using nanopore sequence (longer sequences) in combination with the Illumina sequence (short sequence) a hole genome sequence is possible.
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