RNA-seq 101

rna-seq

Workflow

1. extraction
2. library prep
3. sequencing $\to$ QC
4. mapping/quantification $\to$ QC
5. DGE with R
6. functional analysis with R

Harvard training

• Library preparation (adding adapters), Experimental planning considerations
• Sequencing
• FastQC quality check
• fastp trimming
• STAR alignment

Splice-aware mapping to genome Counting reads associated with genes Statistical analysis to identify differentially expressed genes

1. Pre-alignment QC
2. Align to the reference human genome
3. Post-alignment QC
4. Visualization
5. Quantify transcripts

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Oligonucleotides is a short set (strand) of of DNA/RNA nucleotides.