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Managing Data Modeling GO Workshop 3-6 August 2010 Managing Data Functional modeling strategy Converting between Database IDs Ensembl Biomart UniProt DAVID AgBase ArrayIDer Arrays examples to work on Types of data sets and modeling Commercial array data – more likely to have ID mapping to support functional modeling. Custom/USDA array data – may need to do your own ID mapping: see examples on workshop page. Proteomics data RNA-Seq data sets – computational pipelines to assign GO (GOanna is limited; contact AgBase). Real-time data or quantitative proteomics data – hypothesis testing. Overview of Functional Modeling Strategy Microarray Ids ArrayIDer Protein/Gene identifiers GOModeler hypothesis testing Pathways and network analysis Ingenuity Pathways Analysis (IPA) Pathway Studio Cytoscape DAVID GO Enrichment analysis GORetriever Genes/Proteins with GO annotations no GO annotations GOanna Ingenuity Pathways Analysis (IPA) Pathway Studio Cytoscape DAVID EasyGO/AgriGO Onto-Express Onto-Express-to-go (OE2GO) GOSlimViewer summarizes GO function Yellow boxes represent AgBase tools Green/Purple boxes are non-AgBase resources Functional Modeling Considerations Should I add my own GO? Should I do GO analysis and pathway analysis and network analysis? use GOSlimViewer to see how much GO is available for your species use GORetriever to see how much GO is available for your dataset different functional modeling methods show different aspects about your data (complementary) is this type of data available for your species (or a close ortholog)? What tools should I use? which tools have data for your species of interest? what type of accessions are accepted? availability (commercial and freely available) structurally and functionally re-annotated a microarray quantified the impact of this re-annotation based on GO annotations & pathways represented on the array tested using a previously published experiment that used this microarray re-annotation allows more comprehensive GO based modeling and improves pathway coverage re-annotation resulted in a different model from previously published research findings Converting accessions Depending on your data set & the tools you use, you are likely to need to convert between database accessions to do your functional modeling. UniProt database – ID mapping tab Ensembl BioMart Online analysis tools: DAVID g:profiler GORetriever ArrayIDer – converts EST accessions for some species (by request) ID Mapping Commercial arrays EST arrays Commercial ID mapping eg. NetAffy Ensembl BioMart Online tools (g:convert, DAVID) ArrayIDer Custom arrays Proteomics UniProt ID Conversion RNA-Seq data Working on your own data: New to GO Your own data set GO browser tutorials to familiarize yourself with the GO learn what GO is available for your species functional grouping to get overview (eg. GOSlimViewer GO enrichment analysis (tools available for your species) Pathway analysis Example data sets available – use as worked examples Working on your own data: New to GO Your own data set only certain database GO browser tutorials to familiarize yourself with Most the GO of these tools (including learn what GOAnalysis) is available for your species Pathways accept functional grouping to get overview (eg. accessions GOSlimViewer needanalysis to convert GO enrichment (tools available for your species) accessions between databases Pathway analysis Example data sets available – use as worked examples Example: ID conversion Ensembl Plant Biomart tool currently limited species, but Ensembl is adding more plants BioMart allows sophisticated querying of genomic data DAVID ID conversion tool UniProt ID conversion allows users to convert IDs and do GO enrichment analysis highly annotated data ArrayIDer links ESTs to public database IDs http://plants.ensembl.org/index.html NOTE: Ensembl is adding new plant species… 1. Ensembl BioMart Clicking on these headings allows you to set up searches. Selecting FILTERS gives you different filtering options: Expand GENE and check “ID list limit” to select a defined list of accessions. Enter your list of accessions. Selecting ATTRIBUTES allows you to choose what information is reported: Check accessions from external databases (UniProt & RefSeq). Clicking on RESULTS will show you the output information. Output can be displayed online and/or downloaded (text, Excel). Selecting FILTERS or ATTRIBUTES will allow you to go back and make changes. Limited to species represented in Ensembl 2. Online analysis tools Database for Annotation, Visualization and Integrated Discovery (DAVID) http://david.abcc.ncifcrf.gov/conversion.jsp This tool works for a wide range of species. Paste in your accession list (You can also upload a file of accessions.) Select accession type. NOTE: If you choose “Note Sure” the tool will try to decide what type of accession you have. Select gene list. Submit list. Select the type of accession you want to convert TO. Any ambiguous IDs are listed for you to decide. 3. UniProt ID Mapping Paste accession list (>1000 may cause errors). COMMENT: Note the difference between UniProt Accessions and UniProt IDs. UniProt accessions are a short string a letters and numerals 6-8 characters long. UniProt IDs have a suffix related to the species name. Eg: Cassava Hydroxynitrilase Accession: P52705 ID: HNL_MANES Select the accession type you have: and the accession type you want to convert to: Click on MAP The mapping link will display a tab separated file that can be displayed in Excel: 4. AgBase: ArrayIDer Maps ESTs to gene/protein accessions. Contact AgBase to request additional species. Upload a list of dbEST accessions or EST names. An email will be sent with a link to the results. Results are formatted as an Excel file. For additional help with database accessions please contact AgBase. Working on your own data: NOTE: Always keep note of what tool you used to do the accession ID mapping/conversion and its version/update/date. Keep a copy of your original IDs and what they mapped to so that you can refer back to this during your modeling. Tutorial 1: ID conversion The AgriGO GO enrichment analysis tool accepts the following inputs for rice: GenBank ID: AAP50233.1 DDBJ ID: BAB11514.1 EMBL ID: CAA18188.1 UniProt ID: Q9LYA9 RefSeq Peptide ID: NP_564434 We will convert a list of Rice Affy IDs to these IDs for use in the AgriGO tool. Arrays: ID Mapping “annotation” file that shows which database accessions the probes were based on array annotation files may include multiple database IDs Commercial arrays – may be updated regularly Custom/Research arrays – not updated as often Always check when the last ID mapping was updated, as this data changes continually Array annotation available: FHCRC chicken 13K GPL2863 Agilent-015068 Chicken Gene Expression Microarray 4x44k GPL8764 Avian Innate Immunity Microarray (AIIM) GPL1461 Affymetrix Chicken Genome Array GPL3213* UIUC Bos taurus 13.2K 70-mer oligoarray GPL2853 Affymetrix Bovine Genome Array GPL2112 Agilent-015354 Bovine Oligo Microarray (4x44K) Equine Whole Genome Oligonucleotide (EWGO) array Array annotation in progress: ARK-Genomics G. gallus 20K v1.0 GPL5480 FHCRC Chicken 13K v2.0 GPL1836 Chicken cDNA DDMET 1700 array version 1.0 GPL3265 Tutorial 1: ID conversion Work through tutorial 1 on the workshop website. Alternatively – work on your own data set during this time, using the tutorial as a guide.