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Comparative
transcriptomics
of fungi
Group Nicotiana
Daan van Vliet, Dou Hu, Joost de Jong, Krista Kokki
Research objective
To study differences in gene expression in
related fungi species
Studies species:
- Reference genome
- RNA reads > 100 bp
- Preferably: Paired-end
- Related species (or at least: single-celled,
eukaryotic)
- Similar conditions
Data Management
- Cleaning and trimming reads: FastQC (< 25)
Under Pipeline:
- Mapping reads: TopHat
- Assemble and quantify transcripts: Cufflinks
- Final transcriptome assembly: Cuffmerge
- Extracting the transcripts: gffread
Data Management
Correlation between expressed gene and transcript
properties
• determine GC content and transcript length
• determine intron length
• codon usage (loop analysis codon by codon;
count frequencies of codons in array )
Validation
• Run scripts on small, example datasets
automating in the pipeline
Data Processing
Module building
• Organize the components of our project
• Separate project in functional blocks
Visualization and Interaction network
• Compare different dataset or same species in
different conditions -- Cuffdiff
• R and Cytoscape to make visualization in
interaction network