Bioinformacs Resources - Genbank · 2016. 4. 29. · BioinfRes SoSe 16 Most Growing Divisions...
Transcript of Bioinformacs Resources - Genbank · 2016. 4. 29. · BioinfRes SoSe 16 Most Growing Divisions...
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Bioinforma)csResources-Genbank-
Lecture&ExercisesProf.B.Rost,Dr.L.Richter,J.Reeb
Ins)tutfürInforma)kI12
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Na)onalCenterforBiotechnologyInforma)on,NCBI
http://nihrecord.nih.gov/newsletters/2013/07_19_2013/images/milestonesPic6.jpg
● firstideasinthemiddleofthe80s
● divisionoftheNa)onalLibraryofMedicine(NLM)insidetheNa)onalIns)tutesofHealth(NIH)
● poli)calmission
● foundedin1988
● DavidLipman
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NCBI’spoli)calmissionasdefinedbythebill:1. design,develop,implement,andmanageautomatedsystems
forthecollec)on,storage,retrieval,analysis,anddissemina)onofknowledgeconcerninghumanmolecularbiology,biochemistry,andgene)cs;
2. performresearchintoadvancedmethodsofcomputer-basedinforma)onprocessingcapableofrepresen)ngandanalyzingthevastnumberofbiologicallyimportantmoleculesandcompounds;
3. enablepersonsengagedinbiotechnologyresearchandmedicalcaretousesystemsdevelopedunderparagraph(1)andmethodsdescribedinparagraph(2);and
4. coordinate,asmuchasisprac)cable,effortstogatherbiotechnologyinforma)ononaninterna)onalbasis.
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SelectedNCBIAccomplishmentsBlastGenBankatNCBI
NCBIwebsite
GenomesOMIM
PubMed
1990
1992
1994
1995
1996
1997
HumanGenomePubMedCentral
EntrezGene/DTDs
NIHPublicAccessGenomeReferenceConsor)um
1000GenomesProject
1999
2000
2003
2005
2007
2008
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NCBIResources● NCBIcurrentlyhostsavastbunchofresourceshap://www.ncbi.nlm.nih.gov/guide/all/
● groupedaccordingtovariouscriteria- metadata,project-centric- methodoriented- topicoriented
● sortedinthesec)ons:databases,downloads,submissions,tools,howtos
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Genbank’sOrigin
● WalterGoad,LosAlamosNa)onalLaboratory
● LosAlamosSequenceDatabase1979
● Crea)onandreleaseofGenBankin1982
● Endof1982:2000sequences
● MovetoNCBIin1992http://www.lanl.gov/science-innovation/features/innovations/images/light/thumbnails/21.jpg
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Minutesfrom20thanniversaryofGenBankin2002
“....AmongthemisamemoonLosAlamosNa)onalLaboratorysta)onerydatedMay9,1980,thatreads:Monday,May12at10:30SteveSimoninvitesyouforcakeandcoffeetocelebrate100,000basesnowintheDNAsequencelibrary.”
takenfromhaps://www.genomeweb.com/genbank-turns-20
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GrowthofGenBankandWGS
-doublingapprox.every18months,diagramforrelease207,Apr.2015-currentversion:release213,Apr.2016:211.423.912.047basesinGenbank,1.452.207.704.949basesinWGS-takenfromhap://www.ncbi.nlm.nih.gov/genbank/sta)s)cs
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GrowthofGenBankandWGS
-currentrelease213:193.739.511sequenceinGenbank,338.922.537sequencesinWGS-takenfromhap://www.ncbi.nlm.nih.gov/genbank/sta)s)cs,release207,Apr.2015
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ReferencesforGenBank● thecurrentcita)onsource:“GenBank”.NucleicAcidsRes.2014Jan;42(Databaseissue):D32-7.doi:10.1093/nar/gkt1030.Epub2013Nov11.
● PMID:24217914● partoftheInterna)onalNucleo)deSequenceDatabaseCollabora)on(INSDC)togetherwithEMBLNucleo)deSequenceDatabase(EMBL-Bank),partoftheEuropeanNucleo)deArchive(ENA)andtheDNADataBankofJapan(DDBJ)
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MostGrowingDivisionsDivision Description Release 197
(8/2013) Annual Increase (%)
WGS* Whole-genome shotgun data 500.420.412.665 62.4.
TSA* Transcriptome shotgun data 8.6333123.935 49.9
PHG Phages 119.812.712 42.5
VRL Viruses 1.757.202.472 22.9
BCT Bacteria 10.281.048.518 21.8
ENV Environmental samples 3.743.277.434 10.9
INV Invertebrates 2.737.140.464 9.8
PAT Patented sequences 13.290.161.247 9.7
PLN Plants 5.963.882.822 8.8
GSS Genome survey sequences 23.726.384.753 8.1
VRT Other vertebrates 3.068.956.026 6.3
MAM Other mammals 911.342.025 5.6
... ... ... ...
TOTAL All GenBank sequences 654.613.333.676 45.1
* not distributed with the release; there specific project server sections
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TopOrganisms(Rel.207)Organism Entries Non-WGS base
pair Homo sapiens 20.921.637 17.714.786.437 Mus musculus 9.727.522 9.995.696.539
Rattus norvegicus 2.193.812 6.526.236.496 Bos taurus 2.227.298 5.410.360.312 Zea mays 4.177.175 5.201.714.457 Sus scrofa 3.297.029 4.895.127.638
Danio rerio 1.727.668 3.133.901.682 Triticum aestivum 1.796.780 1.927.718.314
... ... ... Oryza sativa
Japonica Group 1.376.410 1.265.556.227
... ... ... Arabidopsis thaliana 2.578.785 1.202.100.008
... ...
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Distribu)onofSequenceFiles(Rel.207)Division Number of Files
BCT 178 CON 317 ENV 81 EST 478 HTG 142 INV 126 PAT 219 PLN 107 TSA 175 VRL 34
Release 207 consists of 2333 text files in total.
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DatabaseFiles
● GenBankcomesinasetofcompressedtextfilesavailableviaFTP
● seejp://jp.ncbi.nih.gov/genbank/gbrel.txt● 2333ASCIIfiles(listedindivisionplusaddi)onallistfiles)intherangeof0.7-520MB
● uncompressed~709GB● eachfileconsistsoftwopor)ons
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DatabaseFiles● Part1:highlyconserveddatabasefileheaders1 10 20 30 40 50 60 70 79 ---------+---------+---------+---------+---------+---------+---------+--------- GBBCT1.SEQ Genetic Sequence Data Bank April 15 2015 NCBI-GenBank Flat File Release 207.0 Bacterial Sequences (Part 1) 51396 loci, 92682287 bases, from 51396 reported sequences ---------+---------+---------+---------+---------+---------+---------+--------- 1 10 20 30 40 50 60 70 79
● Part1:sequenceentriesforthatdivisiondescribedintheheader
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TheGenBankFlatFileFormat
● asequenceentryconsistsofmanyrecords(lines)● eachrecordconsistsoftwoparts
● Part1:columns1-10/EntryFieldName
● Part2:remaininglinewiththecontent
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Part1/1● akeyword,beginningincolumn1oftherecord(e.g.,REFERENCEisakeyword)
● asubkeywordbeginningincolumn3,withcolumns1and2blank(e.g.,AUTHORSisasubkeywordofREFERENCE)
● orasubkeywordbeginningincolumn4,withcolumns1,2,and3blank(e.g.,PUBMEDisasubkeywordofREFERENCE)
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Part1/2
● blankcharacters,indica)ngthatthisrecordisacon)nua)onoftheinforma)onunderthekeywordorsubkeywordaboveit
● acode,beginningincolumn6,indica)ngthenatureofanentry(featurekey)intheFEATUREStable
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Part1/3● anumber,endingincolumn9oftherecord:- Thisnumberoccursinthepor)onoftheentrydescribingtheactualnucleo)desequenceanddesignatesthenumberingofsequenceposi)ons
● twoslashes(//)inposi)ons1and2,markingtheendofanentry
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Part2● Thesecondpartofeachsequenceentryrecordcontainstheinforma)onappropriatetoitskeyword
● inposi)ons13to80forkeywords
● inposi)ons11to80forthesequence
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EntryFieldTypes(incomplete)● Locus:Ashortmnemonicnamefortheentry,chosentosuggestthesequence'sdefini)on;mandatorykeyword/exactlyonerecord.
● Defini4on:Aconcisedescrip)onofthesequence;mandatorykeyword/oneormorerecords
● Accession:- theprimaryaccessionnumberisaunique,unchangingiden4fierassignedtoeachGenBanksequencerecord.
- tobeusedforcita)onsfromGenBank- mandatorykeyword/oneormorerecords.
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EntryFieldTypes(incomplete)
● Version:- compoundiden)fierconsis)ngoftheprimaryaccessionnumberandanumericversionnumberassociatedwiththecurrentversionofthesequencedataintherecord
- op)onallyfollowedbyanintegeriden)fier(a"GI")assignedtothesequencebyNCBI
- mandatorykeyword/exactlyonerecord
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EntryFieldTypes(incomplete)
● DBLINK:providescross-referencestoresourcesthatsupporttheexistenceasequencerecord;op4onalkeyword/oneormorerecords
● Keywords:shortphrasesdescribinggeneproductsandotherinforma)onaboutanentry;mandatorykeywordinallannotatedentries/oneormorerecords
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EntryFieldTypes(incomplete)
● Source:Commonnameoftheorganismorthenamemostfrequentlyusedintheliterature;mandatorykeywordinallannotatedentries/oneormorerecords/includesonesubkeyword
● Organism:Formalscien)ficnameoftheorganism(firstline)andtaxonomicclassifica)onlevels(secondandsubsequentlines);mandatorysubkeywordinallannotatedentries/twoormorerecords
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EntryFieldTypes(incomplete)● Reference:- Cita)onsforallar)clescontainingdatareportedinthisentry
- includessevensubkeywordsandmayrepeat- mandatorykeyword/oneormorerecords
● Journal:liststhejournalname,volume,year,andpagenumbersofthecita)on;mandatorysubkeyword/oneormorerecords
● op)onalsubkeywords:Authors,Consor)um,Title,Medline,Pubmed,Remark
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EntryFieldTypes(incomplete)● Features:tablecontaininginforma)ononpor)onsofthesequencethatcodeforproteinsandRNAmolecules;sitesofbiologicalsignificance;op4onalkeyword/oneormorerecords
● Origin:- specifica)onofhowthefirstbaseofthereportedsequenceisopera)onallylocatedwithinthegenome
- mandatorykeyword/exactlyonerecord- followedbysequencedata(mul)plerecords)
● //:entrytermina)onsymbol;mandatoryattheendofanentry/exactlyonerecord
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DetailedLocusFormatColumns Contents 01-05 'LOCUS'
06-12 spaces
13-28 Locus name
29-29 space
30-40 Length of sequence, right-justified
41-41 space
42-43 bp
44-44 space
45-47 spaces, ss- (single-stranded), ds- (double-stranded), or ms- (mixed-stranded)
48-53 NA, DNA, RNA, tRNA (transfer RNA), rRNA (ribosomal RNA), mRNA (messenger RNA), uRNA (small nuclear RNA), left justified
54-55 space
56-63 'linear' followed by two spaces, or 'circular'
64-64 space
65-67 The division code
68-68 space
69-79 Date, in the form dd-MMM-yyyy (e.g., 15-MAR-1991)
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AccessionFormat● sixoreightcharacters● sixcharacterformat:- singleuppercaseleaer- 5digits
● eigthcharacterformat:- twouppercaseleaers- 6digits
● primaryaccessionnumberalwaysthefirstone
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Features(Incomplete)
● authorita)vesource:hap://www.insdc.org/documents/feature-table
● featuretablecontainsinforma)onabout:- geneandgeneproducts- regionsofbiologicalsignificance- canenumeratedifferencesbetweenvariousreports- providescross-referencestootherdatacollec)ons- allowshierarchicalrela)onbetweenthefeatures
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Layout● firstlineofthefeaturetableisaheader● includesthekeyword‘FEATURES’andthecolumnheader‘Loca)on/Qualifiers’
● eachfeatureconsistsof:- descriptorlinecontainingafeaturekeyandaloca)on
- acon)nua)onlinefortheloca)onmayfollow- featurequalifiersmayfollowthedescriptorline- key:column6-20,loca)onstartsincolumn22- qualifiersonsubsequentlinesatcolumn22star)ngwitha‘/’
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AFewFrequentFeatures● CDS:sequencecodingforaminoacidsinprotein(includesstopcodon)
● exon:regionthatcodesforpartofsplicedmRNA● gene:regionthatdefinesafunc)onalgene,possiblyincludingupstream(promotor,enhancer,etc)anddownstreamcontrolelements,andforwhichanamehasbeenassigned
● mRNA:messengerRNA
● .......>60featurescurrently
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Loca)onandQualifiers
● Loca)on:- aloca)oncanbe:asinglebase,aspanofbases,asitebetweentwobases,ajoinofsequences,...
- examples:23,23..56,23^24,join(23..56,87..110)
● Qualifiers:- format:fromcolumn22/qualifier_name[=value]- types:freetext,enumera)onorcontrolledvocabulary,cita)ons,sequences,featurelabels
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DatabaseCrossReferences/db_xref
● hap://www.ncbi.nlm.nih.gov/genbank/collab/db_xref/
● Qualifier:/db_xref="database:idenDfier”● Defini4on:databasecross-reference:pointertorelatedinforma)oninanotherdatabase
● Scope:allfeaturekeys● Example:/db_xref="Swiss-Prot:P12345”
● currently>120databasesavailable
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AnatomyofaGenbankFlatFile
. . .
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AnatomyofaGenbankFlatFile
. . .
Locus line
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AnatomyofaGenbankFlatFile
. . . Accession Number, Version and GI number
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AnatomyofaGenbankFlatFile
. . . Feature table with annotations
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UsefulResourcesfromNCBI
● Materials:● Electronicbookshelf
● hap://www.ncbi.nlm.nih.gov/educa)on/factsheets/
● jp://jp.ncbi.nih.gov/pub/factsheets/Factsheet_Books.pdf
● NCBImanuals
● textbooks
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UsefulResourcesfromNCBI
● Processes,e.g.Prokaryo)cGenomeAnnota)onPipeline
● designedforbacterialandarchaealgenomes● mul)-levelprocessincludingprotein-codinggenepredic)onandfunc)onalgenomeunitlikerRNAs,tRNAs,smallRNAs,pseudogenescontrolregions,repeats,inser)onelementsa.s.f.
● combina)onofab-iniDopredic)onandhomologybasedmethods
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UsefulResourcesfromNCBI● referencedatabases:RefSeq● hap://www.ncbi.nlm.nih.gov/refseq/
● comprehensive,integrated,non-redundant,well-annotatedsetofsequences,includinggenomicDNA,transcripts,andproteins
● stablereferenceforgenomeannota)on,esp.subsetofRefSeqGene
● referencesequences
● referencecoordinates● accessibleviaBLAST,EntrezandFTP
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RefSeq● createdby:- Eukaryo)cGenomeAnnota)onPipeline- Prokaryo)cGenomeAnnota)onPipeline- Manualcura)on- SubmissiontoINSDCmembers
● reflectcurrentknowledgeofsequencesdataandbiology
● formatconsistency● Accessionnumbercontainsan“_”
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RefSeqGrowth
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DatabasesAccessibleviaEntrez
http://www.ncbi.nlm.nih.gov/gquery/
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Computa)on:BlastatNCBI
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SearchingtheNCBI/Entrez● provideanintegratedsearchinterfacetothedifferentNCBIdatabases:EntrezProgrammingU)li)es(E-u)li)es)
● Base-URL:hap://eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/
● >40databases
● stableinterfaceofnineserver-sideprograms
● hap://www.ncbi.nlm.nih.gov/books/NBK25501/
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EntrezGuidelines● ifyouusetheeu)lsagainsttheguidelinesyoumightbebanned!
● >100requests:weekendsoroutsideUSpeak)mes(9pm-5am,EST)
● notmorethan3requestpersecond
● provideemailandtoolname:&tool=&email=!
● registra)onwithemailandtoolnamewithNCBImayrelaxtheserestric)ons
● supportedbyBioPython
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Construc)ngURLs
● parameter:&lowerCaseName● excep)on:&WebEnv
● norequiredorder
● nullvaluesandinappropriateparameteraregenerallyignored
● nospaces,use+instead
● useURLencodingsforspecialcharacterlike:%22for“or%23for#or%40for@
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E-u)li)es● Einfo● Esearch
● EPost
● ESummary● EFetch
● ELink
● EGQuery
● ESpell● ECitMatch
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ESearch
● textsearch● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/esearch.fcgi
● respondstoatextquerywiththelistofmatchingUIDsinagivendatabase(forlateruseinESummary,EFetchorELink),alongwiththetermtransla)onsofthequery
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ESummary
● documentsummarydownloads● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/esummary.fcgi
● respondstoalistofUIDsfromagivendatabasewiththecorrespondingdocumentsummaries
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EGQuery
● globalquery● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/egquery.fcgi
● respondstoatextquerywiththenumberofrecordsmatchingthequeryineachEntrezdatabase
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EInfo
● databasesta)s)cs● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/einfo.fcgi
● providesthenumberofrecordsindexedineachfieldofagivendatabase,thedateofthelastupdateofthedatabase,andtheavailablelinksfromthedatabasetootherEntrezdatabases
● without&db:listsallavailabledatabases
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EFetch
● datarecorddownloads● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/efetch.fcgi
● respondstoalistofUIDsinagivendatabasewiththecorrespondingdatarecordsinaspecifiedformat
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ELink
● Entrezlinks● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/elink.fcgi
● respondstoalistofUIDsinagivendatabasewitheitheralistofrelatedUIDs(andrelevancyscores)inthesamedatabaseoralistoflinkedUIDsinanotherEntrezdatabase
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ELink
● checksfortheexistenceofaspecifiedlinkfromalistofoneormoreUIDs
● createsahyperlinktotheprimaryLinkOutproviderforaspecificUIDanddatabase,orlistsLinkOutURLsandaaributesformul)pleUIDs
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EPost
● UIDuploads● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/epost.fcgi
● acceptsalistofUIDsfromagivendatabase,storesthesetontheHistoryServer,andrespondswithaquerykeyandwebenvironmentfortheuploadeddataset
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ESpell
● spellingsugges)ons● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/espell.fcgi
● retrievesspellingsugges)onsforatextqueryinagivendatabase
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ECitMatch
● batchcita)onsearchinginPubMed● eu)ls.ncbi.nlm.nih.gov/entrez/eu)ls/ecitmatch.cgi
● retrievesPubMedIDs(PMIDs)correspondingtoasetofinputcita)onstrings
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Iden)ficators● recordsareiden)fiedbyanintegerIDcalledUID● UIDaredatabasespecificlikeGInumbers,PMIDS,MMDB-IDs
● UIDareaswellinputandoutput
● especiallyusefulincombina)onwiththeHistoryserver
● afulldescrip)onofparametersandsyntaxcanbefoundat:hap://www.ncbi.nlm.nih.gov/books/NBK25499/
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SelectedUIDsEntrez Database UID common name E-utility Database Name Books Book ID books Conserved Domains PSSM-ID cdd dbVar dbVar ID dbvar EST GI number nucest Gene Gene ID gene Genome Genome ID genome MeSH MeSH ID mesh NCBI Web Site Web Site ID ncbisearch Nucleotide GI number nuccore PubMed PMID pubmed ... ... ...
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EntrezCoreEngine● EGQuery,ESearch,andESummary● twotasks:- assemblealistofUIDsthatmatchatextquery(ESearch)- retrieveabriefsummaryrecordcalledaDocumentSummary(DocSum)foreachUIDESummary)
● EGQuey:globalversionofESearch● esearch.fcgi?db=database&term=query esummary.fcgi?db=database&id=uid1,uid2,uid3,...!
● expandedintomorecomplicatedEntrezqueries
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EntrezDatabases(EInfo,EFetch,andELink)
● EInfo:- providesdetailedinforma)onabouteachdatabase- includinglistsoftheindexingfieldsinthedatabase- availablelinkstootherEntrezdatabases
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EntrezDatabases(EInfo,EFetch,andELink)
● addedvaluetotherawdata:- supportsavarietyofdisplayformats:EFetchUIDlistsinXMLandplaintext(&retmode)foralldatabases,otherformats(&rettype)aredatabasespecific
- hap://www.ncbi.nlm.nih.gov/books/NBK25499/table/chapter4.T._valid_values_of__retmode_and/?report=objectonly
- efetch.fcgi?db=database&id=uid1,uid2,uid3 &rettype=report_type&retmode=data_mode!
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EntrezDatabases(EInfo,EFetch,andELink)
● addedvaluetotherawdata:- linkstorecordsinotherEntrezdatabasesmanifestedaslistofassociatedUIDs
- UIDsmustbevalidinsourcedatabase(&dbfrom)- elink.fcgi?dbfrom=protein&db=gene&id=15718680,157427902
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EntrezHistoryServer
● simple:intheGUIaccessibleviatherespec)vetabs
● youcanstoretemporarilysetsofUIDsasinputforlaterqueriesthroughothertools
● eachlistofUIDsisspecifiedby:- &query_key(integerlabel)- &WebEnv(cookiestring)
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Crea)onofastoredUIDlist
● EPost:- EPostcanbeuseduploadaUIDlist- returns&query_keyand&WebEnv!
● ESearch:- storestheresultsifgiven&usehistory=y!
● ELink:- storestheresultsifgiven&cmd=neighbor_history!
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UsageofstoredUIDlists● Useofstoredlists:esummary.fcgi?db=database&WebEnv=webenv &query_key=key!
● onewebenvironmentcanholdmul)pleresultlists
● listsinthesamewebenvironmentcanbecombinedwithAND,OR,NOT
● bydefaulteverycallcreatesanewenvironment
● ->give&WebEnvinsubsequentcallstostorethelistsinthesamewebenvironment
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SketchingPipelines
● getDocSummariesorentriesforkeywordsorIDs:- ESearch->ESummary/EFetch- EPost->ESummary/EFetch
● filter/limitarecordset:- EPost/ELink->ESearch
● moreadvancedqueries:- ESearch->ELink->ESummary/EFetch- EPost->ELink->ESearch->EFetch
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E-u)lityWebinar
● haps://www.youtube.com/watch?v=iCFVVexp30o