Before we start •Login to the laptop: •user: crgcomu •Password: crgcomu •Login to the network: •Wifi: carretwifi •Password : ana@bong •Login to galaxy (ldap): •User: your web mail user •Password : your web mail password Galaxy: an Open Web-Based Analysis Platform Jean-François Taly & Ernesto Lowy Bioinformatics Unit CRG (Barcelona, Spain) Introduction Developed by Nekrutenko and others at Penn State, along with James Taylor at Emory University Galaxy is an open web-based tool for biomedical research Accessible: users without programming experience can easily specify parameters and run tools and workflows Reproducible: Galaxy captures information so that any user can repeat and understand a complete computational analysis Galaxy Interface Input Output history Browsing Data Running Tool: Bowtie2 28 Public Galaxy Servers Galaxy Tutorials Galaxy Workflows Community Tools Community Tools Hardware schema Galaxy: How to Upload Data? Jean-François Taly & Ernesto Lowy Bioinformatics Unit CRG (Barcelona, Spain) Galaxy Interface Input Output history Get Data From UCSC Get Data From UCSC Get Data From BioMart Get Data From BioMart Upload Local Data Upload Local Data From your disk From web server From ftp server Upload Data: From Disk From your disk Upload Data: File Format Upload Data: From Disk Upload Data: Genome Upload Data: From Disk Purple = upload is being prepared Upload Data: From Disk Yellow = upload is running Upload Data: From Disk Green = upload is finished Browsing Data Edit Data Attributes Upload Data: FTP FTP must be used for big files (>2Gb) Process in 2 steps: 1. Use a FTP client to TEMPORARILY upload the file into the server 2. Use Galaxy to move the file to its final destination Time before deletion: ◦ ◦ In step 1 : 2 weeks In step 2 : Forever Upload Data: FTP FTP server name: galaxy.crg.es Username and Password are the same as the ones of Galaxy! You need a FTP client: ◦ ◦ ◦ ◦ Filezilla (all platforms) WinSCP (for Windows) Fetch (for MAC) gFTP (for Linux) Upload Data: FTP (Filezilla) Upload Data: FTP (Filezilla) Upload Data: FTP (Filezilla) Do it yourself! Task: 1.Use your prefered FTP client to upload the file “Human-SE-RNAseq.fastq” in the FTP server Tip: 1.Server name: galaxy.crg.es 2.Username & Password: Same as for your CRG mail Upload Local Data Upload Local Data Galaxy: ChipSeq Jean-François Taly & Ernesto Lowy Bioinformatics Unit CRG (Barcelona, Spain) Chip Seq Goal of exercice Input is Chip Seq Reads from ◦ Specie: mouse, chr19 ◦ Cell line: G1E_ER4 Conditions: with or without: ◦ Transcriptional repressor CTCF Align reads to the reference genome Use the treated and control alignments to find peaks Do it yourself! Task: 1.Use the method you want to upload the file “G1E_ER4_CTCF_chr9.fastqsanger” in your history Tip: 1.The file is the repertory “experiment1” you should have copied from the pen drive. 2.Do not forget the datatype Input Data Quality Check Quality Check Running Tool: Bowtie2 Load a file while computing Do it yourself! Task: 1.Use the method you want to upload the file “” in your history 2.Map the reads to the genome mm9 with bowtie2 Tip: 1.The file is the repertory “” you should have copied from the pen drive. Prepare peak calling (MACS) Prepare peak calling (MACS) Check the parameters Genome Browser: Trackster Trackster: Create New Trackster: Create New Trackster Trackster: Add track Trackster: Add track Workflow Workflow Workflow Workflow Do it yourself! Task: 1.Use the method you want to upload the files “G1E_CTCF.fastqsanger” and “G1E_input.fastqsanger” in your history Tip: 1.The file is the repertory “experiment2” 2.Do not forget the datatype 3.First the CTCF file and then the control Workflow Workflow Workflow