1.2 Expression Matrix
Pipeline
Data Structure
Inputs
File format | Information contained in file | File description | Notes |
bam | alignments | Produced by mapping reads to the transcriptome. | Reads are trimmed using a proprietary version of cutAdapt. We map to transcriptome for a better sensitivity (see details in protocol and example). |
Outputs
File format | Information contained in file | File description | Notes |
bigWig | signal | Normalized RNA-seq signal | Signals are generated for transcriptome both the plus and minus strands and for unique reads and unique+multimapping reads. |
tsv | gene (ncRNA) quantifications | Non-normalized counts. |
Running Scripts
Software/Tools
RSEM
homer
HTseq
FeatureCounts
Example of single case
Tips/Utilities
Merge multiple bams to one
Homework and more
Visualize your mapped reads with IGV (locally) and/or UCSC Genome Browser (on line).
Learn how to construct the expression matrix using HTSeq, featureCounts and homer; then compare the difference among these three methods.
More Reading and Practice
Additional Tutorial : 2. Construction of expression matrix
Bioinformatics Data Skills
11) Working with Alignment Data
Video
a) Expression matrix
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