R / Bioconductor for Sequence Analysis Martin Morgan 1 June 20-23, - PowerPoint PPT Presentation

R / Bioconductor for Sequence Analysis Martin Morgan 1 June 20-23, 2011 1 mtmorgan@fhcrc.org

Bioconductor Goal Help biologists understand their data ◮ Expression and other microarray Focus ◮ Sequence analysis ◮ Imaging, flow cytometry, . . . ◮ Based on the R programming language – Themes statistics, visualization, interoperability ◮ Reproducible – scripts, vignettes , packages ◮ Open source / open development ◮ Contributions from ‘core’ members and (primarily academic) user community Status > 460 packages; very active web site and mailing list; annual conferences; courses; . . .

Using R / Bioconductor Flexibility ◮ Programming language Leveraging resources, e.g., > library(GEOquery) SQL, XML, third party > eset = getGEO( ' ... ' ) libraries (e.g., samtools ) ◮ Scripts, vignettes, packages R statistical methods and ◮ Appeal visualization

Using R / Bioconductor ◮ Programming language 1. Reproducibility > library(GEOquery) > eset = getGEO( ' ... ' ) 2. Communication ◮ Scripts, vignettes, packages 3. Enabling ◮ Appeal

Using R / Bioconductor ◮ Programming language Statisticians > library(GEOquery) > eset = getGEO( ' ... ' ) Bioinformaticists ◮ Scripts, vignettes, packages . . . but not everyone! ◮ Appeal

A Package Tour Bioconductor Pre-processing ◮ Expression and other microarrays Quality assessment ◮ Sequence analysis Differential expression (e.g., limma ) ◮ Annotation and archive Gene set enrichment resources ◮ Additional Many features for free, e.g., machine learning, visualization All of CRAN

A Package Tour Bioconductor ◮ Expression and other microarrays Array CGH (e.g., DNAcopy ) ◮ Sequence analysis Methylation, epigenetics, miRNA ◮ Annotation and archive resources Genotyping (e.g., snpStats ) ◮ Additional All of CRAN

A Package Tour Bioconductor ◮ Expression and other I/O, QA, manipulation microarrays RNAseq differential representation ◮ Sequence analysis (e.g., DESeq ) ◮ Annotation and archive Gene set analysis (e.g., goseq ) resources ChIPseq ◮ Additional Metabiome All of CRAN

A Package Tour 50 ovarian cancer, 13 benign / normal RNAseq samples Bioconductor ◮ Expression and other microarrays ◮ Sequence analysis ◮ Annotation and archive resources ◮ Additional All of CRAN

A Package Tour Differential representation in SOC vs. Control Bioconductor ◮ Expression and other microarrays ◮ Sequence analysis ◮ Annotation and archive resources ◮ Additional All of CRAN

A Package Tour Bioconductor KEGG terms under-represented in ◮ Expression and other SOC microarrays Description P Value ◮ Sequence analysis 1 Spliceosome 0.0017 ◮ Annotation and archive 3 Ribosome 0.0073 resources 5 Cell cycle 0.0123 ◮ Additional ... All of CRAN Investigate intron abundances

A Package Tour Curated, versioned (semi-annual) Bioconductor ◮ Chip ◮ Expression and other microarrays ◮ Organism ◮ Sequence analysis ◮ Pathway ◮ Annotation and archive ◮ Homology resources ◮ miRNA ◮ Additional biomaRt , UCSC All of CRAN GEO , ArrayExpress , SRA

A Package Tour Examples: Identify human genes in Bioconductor ‘spliceosome’, ‘ribosome’, and ‘cell ◮ Expression and other cycle’ KEGG pathways. microarrays Discover and retrieve GEO ◮ Sequence analysis expression arrays related to ovarian ◮ Annotation and archive carcinomas. resources Remotely query 1000 genomes BAM ◮ Additional files for regions of interest, e.g., All of CRAN ‘spliceosome’ genes. Input TCGA ovarian cancer copy number and clinical data.

A Package Tour 86 Paired HMS HG-CGH-244A TCGA samples Bioconductor ◮ Expression and other microarrays ◮ Sequence analysis ◮ Annotation and archive resources ◮ Additional All of CRAN

A Package Tour Bioconductor ◮ Expression and other microarrays Pathways and networks ◮ Sequence analysis Flow cytometry ◮ Annotation and archive High-throughput qPCR resources Image processing ( e.g., EBImage ) ◮ Additional All of CRAN

A Package Tour Bioconductor ◮ Expression and other microarrays 3000+ packages ◮ Sequence analysis Novel approaches, e.g., cghFLasso ◮ Annotation and archive Advanced statistical analyses, e.g., resources Bayesian network models ◮ Additional All of CRAN

Common work flows Input / output ◮ Fasta, fastq – ShortRead ◮ SAM / BAM, tabix, indexed fasta – Rsamtools ◮ Genome tracks & related formats – rtracklayer Pre-processing / manipulation / count & measure ◮ String manipulation, pattern matching Biostrings ◮ Quality assessment ShortRead ◮ finding / counting overlaps GenomicRanges Analysis domains ◮ RNAseq, e.g., DESeq , edgeR , goseq ◮ ChIPseq, e.g., ChIPpeakAnno Annotation / variants ◮ AnnotationDbi / org.* , GenomicFeatures , BSgenome , biomaRt

Useful data structures Biostrings , BSgenome ◮ XString , XStringSet GenomicRanges ◮ GappedAlignments – CIGAR ◮ GRanges / GRangesList – sequence, strand IRanges ◮ IRanges / IRangesList / RangedData – ranges ◮ Rle – run length encoding ◮ Views

Effective compulational software Effective computational biology software 1. Extensive: data, annotation 2. Statistical: volume, technology, experimental design 3. Reproducible: long-term, multi-participant science 4. Current: novel, technology-driven 5. Accessible: affordable, transparent, usable

Bioconductor Who ◮ FHCRC: Herv´ e Pag` es, Marc Carlson, Nishant Gopalakrishnan, Valerie Obenchain, Dan Tenenbaum, Chao-Jen Wong ◮ Robert Gentleman (Genentech), Vince Carey (Harvard / Brigham & Women’s), Rafael Irizzary (Johns Hopkins), Wolfgang Huber (EBI, Hiedelberg) ◮ A large number of contributors, world-wide Resources ◮ http://bioconductor.org: installation, packages, work flows, courses, events ◮ Mailing list: friendly prompt help ◮ Conference: Morning talks, afternoon workshops, evening social. 28-29 July, Seattle, WA. Developer Day July 27