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de.NBI - CeBiTec Nanopore Workshop 2020 - Best Practice and SARS-CoV-2 applications - Overview course structure

Day 1

Start virtual machines

Live demo together with participants

Connect to virtual machines

Linux Introduction

Readthedocs??

Day 2: Basecalling and ReadQC on ARTIC daaset

Connect to Vm and create workdir link

Get data and inspect

  • Download our ARTIC dataset (full dataset and smaller dataset)
  • Download Wuhan Reference (+Annotation!)
  • Exercise?: Inspect the raw data using h5tools

Basecalling

  • (optional): Start Rampart
  • Exercise: Start the basecalling with the smaller dataset
  • (optional): Watch progress in Rampart
  • Inspect the results
  • (optional): Rampart for full dataset
  • Start the basecalling with complete ARTIC dataset (or download finished basecalling)
  • Exercise: Demultiplex

Read QC ARTIC dataset

  • (optional) Use MinIONQC on ARTIC dataset
  • Exercise: Use FastQC on ARTIC dataset
  • Exercise: Use Porechop on ARTIC dataset
  • (optional) Exercise: AlignQC with ARTIC dataset
  • Exercise: Mapping of reads to Wuhan Reference
  • Inspect with IGV/Genomeview

Day 3

Assembly of ARTIC dataset

  • Exercise: start canu with ARTIC dataset
  • Inspect results
  • (optional) Exercise: Map contigs to reference and inspect in IGV/Genomeview
  • Notice, that it didn't work well... -> use other dataset for assembly

Read QC WGS dataset

  • Download WGS dataset
  • Create subsample (or provide it)
  • Exercise: Use FastQC on WGS dataset
  • Exercise: Use Porechop on WGS dataset
  • Exercise: Map WGS reads to reference and inspect in IGV/Genomeview

Assembly of WGS dataset

  • Exercise: Assembly with canu
  • Exercise: Assembly evaluation with quast
  • Exercise: Quality control by mapping

Polishing

  • Exercise: Polish with racon/minimap2 and medaka
  • Exercise: Assembly evaluation with quast
  • Hint for pilon, when Illumina data is available
  • (optional) Exercise: Polish with nanopolish and evaluate with quast

Day 4

ARTIC dataset

  • Exercise: Read filtering
  • Exercise: Run ARTIC pipeline (medaka and nanopolish?)
  • Exercise: Run ARTIC pipeline for all barcodes
  • Evaluation: quast? IGV? ...
  • Load results in nextstrain / phylogenetic tree

Bonus?

Provide old course data and Exercises?