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Tutorial for BIO302_Group-Project-B

Students: Jeanine & Jan

General questions for your project

What does it take from sequencing to a reference genome?

  • address different sequencing techniques, the value and weekpoints of them
  • different assemblers (why so many of them??)
  • discuss the quality of a reference genome assembly (what are the gold standards and what is 'acceptable' to address genomics-related questions)

While focusing on these type of questions, review the avaialbe methods/software, explain how they work and results that you get from them

The projects are designed to cover the various topics of the lectures. With LAB projects, the goal is that students learn each topic in more depth from the students who are working on the specific project in that topic

Study system

Primula grandis (Primulaceae) Endemic to Caucasus; Homostylous (only one flower morph, due to loss of heterostyly)

  • Genome size: 900Mb (Flow Cytometry and K-mer count)
  • Ploidy: tetraploid, 2n=44
  • Data generated: Nanopore (35x total), WGS shotgun, Hi-C (Arima)

Primula grandis

On this Galaxy history, you find the following data

  • Whole Genome Shotgun Sequencing of Primula grandis. This is Illumina short reads, paired-end (PE), 150bp.
  • Nanopore reads of P. grandis

& Narcis will add the following:

  • Hi-C data. This is also Illumina short reads, paired-end (PE), 150bp, but the read1 and read2 come from stretches of DNA which are in close distance in 3D. To refresh your knowledge on Hi-C data, please have a look at LEC2, NGS data generation and handling.
  • Reference assembly of P. grandis generated using both long reads (Nanopore) and short reads (Illumina), which will be used for comparison with your de novo assembly

The project will be done in four steps:

  • genome profiling using GenomeScope2
  • assembly using ABySS (for short reads) and Shasta (for long reads)
  • scaffolding using YaHS
  • annotation using Repeat Modeler (and Maker if we have time)

You find tutorials for genome profiling, assembly, scaffolding and annotation in this repository. Just navigate to different directories.

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