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UCSC Genome Browser 1 The Progress Data from NCBI and TIGR (www.ncbi.nlm.nih.gov and www.tigr.org ) Today - 85,759,586,764 Official “15 year” Human Genome Project: 1990-2003 2 Database and Tool Explosion The annual database issue of Nucleic Acids Research has grown exponentially 1996: first annual compilation of databases and tools lists 57 databases and tools 2008: 1078 databases and tools listed in compilation 2000: 230 databases and tools listed in compilation 3 Genome Browsers UCSC Genome Browser EBI Ensembl NCBI Map Viewer 4 Organizing the Genome Gene X genes & predictions variations and repeats Description Transcript data Structure Gene Ontology Pathway Data Homologous Genes Expression Data Etc…. cross-species comparative data and many more types of data from expression and regulation to mRNA and ESTs… 5 UCSC Genome Browser: genome.ucsc.edu UCSC Genome Browser 7 Organization of genomic data… Annotation Tracks sequence Genome backbone: base position number chromosome band sts sites gap locations known genes predicted genes Links out to more data microarray/expression data evolutionary conservation SNPs repeated regions more… 8 gene details Annotation Tracks official sequence comparisons SNPs A sample of what we will find: 9 The Genome Browser Gateway start page, basic search text/ID searches Use this Gateway to search by: ◦ ◦ ◦ ◦ Gene names, symbols Chromosome number: chr7, or region: chr11:1038475-1075482 Keywords: kinase, receptor IDs: NP, NM, OMIM, and more… See lower part of page for help with format 10 The Genome Browser Gateway start page choices, December 2006 1 2 3 4 5 6 Make your Gateway choices: 1. Select Clade 2. Select species: search 1 species at a time 3. Assembly: the official backbone DNA sequence 4. Position: location in the genome to examine 5. Image width: how many pixels in display window; 5000 max 6. Configure: make fonts bigger + other choices 11 The Genome Browser Gateway sample search for Human TP53 Sample search: human, March 2006 assembly, tp53 select Select from results list ID search may go right to a viewer page, if unique 12 Overview of the whole Genome Browser page } Genome viewer section Groups of data Mapping and Sequencing Tracks Genes and Gene Prediction Tracks mRNA and EST Tracks Expression and Regulation Comparative Genomics Variation and Repeats 13 Sample Genome Viewer image, TP53 region base position STS markers Known genes RefSeq genes GenBank seqs 17 species compared single species compared SNPs repeats 14 Visual Cues on the Genome Browser Tick marks; a single location (STS, SNP) 3' UTR exon <<< exon < exon < < < <ex 5' UTR Intron, and direction of transcription <<< or >>> Track colors may have meaning—for example, Known Gene track: •If there is a corresponding PDB entry, = black •If there is a corresponding NCBI Reviewed seq, = dark blue •If there is a corresponding NCBI Provisional seq, = light blue For some tracks, the height of a bar is increased likelihood of an evolutionary relationship (conservation track) 15 Options for Changing Images: Upper Section Walk left or right click to zoom 3x and re-center Zoom in Specify a position Zoom out fonts, window, more Change your view or location with controls at the top Use “base” to get right down to the nucleotides Configure: to change font, window size, more… 16 Annotation Track display options enforce changes Links to info and/or filters Some data is ON or OFF by default Change track view Menu links to info about the tracks: content, methods You change the view with pulldown menus After making changes, REFRESH to enforce the change 17 Annotation Track options, defined Hide: removes a track from view Dense: all items collapsed into a single line Squish: each item = separate line, but 50% height + packed Pack: each item separate, but efficiently stacked (full height) Full: each item on separate line 18 Reset, Hide, Configure or Refresh to change settings enforce any changes (hide, full, squish…) reset, back to defaults start from scratch You control the views Use pulldown menus Configure options page 19 Click Any Viewer Object for Details Click the item New web page opens Example: click your mouse anywhere on the TP53 line Many details and links to more data about TP53 20 informative description other resource links Click annotation track item for details pages Not all genes have links to sequences This much detail. microarray data Different annotation tracks carry different data. mRNA secondary structure protein domains/structure homologs in other species Gene Ontology™ descriptions mRNA descriptions pathways 21 Get DNA, with Extended Case/Color Options Use the DNA link at the top Plain or Extended options Change colors, fonts, etc. 22 Get Sequence from Details Pages Click a track, go to Sequence section of details page Click the line Click the item sequence section on detail page 23 Accessing the BLAT tool BLAT = BLAST-like Alignment Tool Rapid searches by INDEXING the entire genome Works best with high similarity matches See documentation and publication for details ◦ Kent, WJ. Genome Res. 2002. 12:656 24 BLAT tool overview: Make choices Paste one or more sequences DNA limit 25000 bases Protein limit 10000 aa 25 total sequences submit Or upload 25 go to alignment detail go to browser/viewer BLAT results, with links sorting Results with demo sequences, settings default; sort = Query, Score ◦ Score is a count of matches—higher number, better match Click browser to go to Genome Browser image location (next slide) Click details to see the alignment to genomic sequence (2nd slide) 26 BLAT results, browser link click to flip frame query From browser click in BLAT results A new line with your Sequence from BLAT Search appears! Watch out for reading frame! Click - - - > to flip frame Base position = full and zoomed in enough to see amino acids 27 BLAT results, alignment details Your query Genomic match, color cues Side-by-side alignment yours genomic 28 Proteome Browser Access from homepage or Known Gene pages Exon diagram, amino acids… Many protein properties (pI, mw, composition, 3D…) more data 29 In-Silico PCR: Find genomic sequence using primers Select genome Enter primers Minimum 15 bases Flip reverse primer? Submit 30