introduction to bioinformatics dna technology...
TRANSCRIPT
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Introduction to bioinformatics
DNA technology course
Barbera van SchaikBioinformatics LaboratoryAcademic Medical Centre (AMC)[email protected]
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Health careMolecular Biology/Genomics
Clinical genomics
Information Management / e-Science
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Lost in translation (biology and informatics)
Protocol
NGS
Infrastructure
String
Sequence
Sonic hedgehog
Inheritance
Virus
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Definitions of bioinformatics
Adapted definition according to Wikipedia
The application of information technology and statistics to the field of molecular biology.
The creation and advancement of databases, algorithms, computational and statistical techniques, and theory to solve formal and practical problems arising from the management, analysis and interpretation of biological data.
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What is bioinformatics?
Extraction of biological knowledge from complex data
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What is bioinformatcs?
... one of the results *might* be a tool you can use
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Bioinformatics
Data Knowledge
*convert data to knowledge*generate new hypotheses
*Design new experiments
Experimental
From publicdatabases
And informaton management.........
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How does molecule A interact with protein B?
A schematic visual model of oxygen-binding process, showing all four monomersand hemes, and protein chains only as diagramatic coils, to facilitate visualization into the molecule. (http://en.wikipedia.org/wiki/Hemoglobin)
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Study migration
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Study evolution
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Which gene(s) causes disease X?
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Subjects inbioinformatcs
Genome analysis
Sequence analysis
Phylogenetcs
Structural bioinformatcs
Gene expressionGenetcs and populaton analysis
Systems biology
Data and text mining
Databases and ontologies
http://www.oxfordjournals.org/our_jjournals/bioinformatcs/for_jauthors/scope_jguidelines.html
Bioimage informatcs
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BioinformatcsGroups and insttutes
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Bioinformatics Laboratory (AMC)Medical Bioinformatics
• Part of the KEBB• You are welcome if you need bioinformatics expertise• www.bioinformaticslaboratory.nl
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Focus
Systems genomics (top-down)
(Statistical) analysis of (multi-)omics data
Systems biology (bottom-up)
Mathematical modelling
Information management
Knowledge bases
Science gateways
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Dutch Techcentre for Life Sciences (DTL)
• Support and Education• http://www.dtls.nl/
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Other bioinformatics organisations
• European Bioinformatics Institute (EBI)– http://www.ebi.ac.uk/
• National Center for Biotechnology Information (NCBI)
– http://www.ncbi.nlm.nih.gov/
• EMBnet– http://www.embnet.org/
• International Society for Computational Biology (ISCB)
– http://www.iscb.org/
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Bioinformatics
Extraction of biological knowledge from complex data
@AMC: genomics
, metabolom
ics
and proteo
mics data
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The application of high-throughput automated technologies to molecular biology.
OR
The experimental study of complete genomes.
What is genomics?
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Sample storage
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DNA microarrays
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High throughput sequencing
Run: 2-4 hrsData: 10 GBReads: 60-80 millionLength: 200bp
Run: 4-55 hoursData: 15 GBReads: 25 millionLength: 2x300bp
Run: 1-3.5 daysData: 1500 GBReads: 5 billionLength: 2x150bp
2005-now: Next generation sequencing Millions to billions of sequences
Ion ProtonIllumina MiSeq
Illumina HiSeq (VUmc)
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DNA sequencing
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Sanger sequencing
From DNA toAutoradiograms
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ABI 3100 Automated Capillary DNA Sequencer
Automated Sequencing
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Electropherogram
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Sequencing factoriesCustom-designed factory-style conveyor belt robots:
perform all functions from purifying DNA from bacterial cultures through setting up and purifying sequencing reactions.
Automated sequencing
Whitehead insttute
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2001 The HGP consortium publishes its
working draft in Nature (15 February), and Celera publishes its draft in Science (16 February).
Human genome: 3 billion bases
In total 23 billion bases were sequenced (7.5-fold coverage)
This comprises 23 Gbyte of data
Public Human Genome project
-1990-2003
- 3 billion US dollar
Private Celera genome project
-1998 - 2001
- Craig Venter
- 300 million US dollar 2001
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Traditional versus high throughputDNA sequencing
Roche 454, one run:
7.5 hours
1,000,000 sequences
750 nt per sequence
35 GB data (including images)
1 GB data (excluding images)
= 750,000,000 bases
Year: 2005 – 2015
http://www.454.com/
Illumina, one run:
1-3.5 days
5 billion sequences
2x 150 nt per sequence
1.5 TB data
Year: 2008 – now
Sanger, one run:
? hours (human genome took 15 years)
1-384 sequences
300-1000 nt per sequence
1 KB - 384 KB data
= 1,000-384,000 bases
Year: 1963 – now
http://www.illumina.comhttp://en.wikipedia.org/
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Data size challenges
COMPUTINGSequencing rate is higher
than Moore's law
STORAGESequencing costs lower
than data storage
Stein, Genome Biology, 2010Hayden, Nature, 2014
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Genome projects
• Human genome project (1 genome)• Exome sequencing (~10 individuals)• Genome of the Netherlands (770 individuals)• 1000 genome project (1000 individuals)• 10K UK project (10,000 individuals)
– Upgraded to 100K genomes
… many centers have one or morehigh throughput sequencers
http://omicsmaps.com/
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image credit: Digital Vision, PhotoDisc, Matt ay/EHP
Sign @ Wellcome-Sanger, Cambridge, UK
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Small cluster
Existing hardware
PC
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Buy a bigger cluster (centralized model)
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Dutch life science grid
grid
https://www.surfsara.nl/
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Cloud computng
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Not only hardware
• Software• Peopleware• Sharing
e-Science
More info:http://www.egi.eu/http://www.ebioscience.amc.nl/
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Findable Accessible Interoperable Reproducable (FAIR)
Wilkinson et al. (2016) Scientific Data
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Tips and tricks… where to start?
Biological databasesPublicly available tools
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Publicly available biological databases
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Molecular Biology Database Collection 2015
• Published by Nucleic Acids Research (scientific journal)• Currently 1552 databases.• Latest issue: 56 new databases• Coverage is far from exhaustive.
?
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Database access
• Web–interface– Keyword search– Browsing / cross-linking– BLAST
• From other applications– Applicaton programming interface (API)– Web services– In-house / third party sofware
• Download– Full database– Specifc records– Diferent formats (text, xml, rdf, etc)
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Sequence analysis
Function prediction (similarity, sequence search)Localisation (genefinding)
Grouping (genes, protein families)Conservation (motifs, functional blocks)
SNPs and mutations (variations)
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Sequence analysis
Pairwise alignment: in-exact matching of 2 sequences
Multple sequence alignment: in-exact matchingof >2 sequences
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Blast output
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Blast output - alignments
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Galaxy
https://galaxyproject.org/
Tip: Has instructon videos for a few use casese.g. for analyzing next gen sequence data
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Nucleic acid research – annual web server issue
https://links.bioinformatcs.ca/
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Subjects inbioinformatcs
Genome analysis
Sequence analysis
Phylogenetcs
Structural bioinformatcs
Gene expressionGenetcs and populaton analysis
Systems biology
Data and text mining
Databases and ontologies
http://www.oxfordjournals.org/our_jjournals/bioinformatcs/for_jauthors/scope_jguidelines.html
Bioimage informatcs
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Topics computer exercises
Query public databasesSequence format conversion
Automatic RNA protein translationPrimer designGene finding
Tomorrow 13:30-15:00
https://btioifoomatpcsatacaf//