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Uhlalutyo lwe-Genome-wide Association
I-Genome-wide association study (GWAS) ijolise ekuchongeni ukuhlukahluka kofuzo (i-genotype) ehambelana neempawu ezithile (phenotype).Uphononongo lwe-GWAS luphanda iimpawu zofuzo ezinqamlezileyo kwi-genome yenani elikhulu labantu kwaye liqikelela imibutho ye-genotype-phenotype ngohlalutyo lwamanani kwinqanaba labantu.Isetyenziswe ngokubanzi kuphando ngezifo zabantu kunye nokwembiwa kwemfuza okusebenzayo kwiimpawu ezinzima zezilwanyana okanye izityalo.
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I-nucleus ye-RNA enye
Ukuqhubela phambili ekuthathweni kweseli enye kunye nobuchule bokwakha ithala leencwadi ngalinye kudityaniswe nolandelelwano olugqithileyo luvumela izifundo zokuchazwa kofuzo ngokwesiseko seseli-nge-seli.Ivumela uhlalutyo olunzulu kunye olupheleleyo lwenkqubo kwiiseli ezintsonkothileyo, apho ubukhulu becala inqanda ukufihla ukungafani kwazo ngokuthatha umndilili wazo zonke iiseli.
Nangona kunjalo, ezinye iiseli azifanelekanga ukuba zenziwe zibe yi-single-cell suspension, ngoko ke ezinye iindlela zokulungiselela iisampulu-i-nucleus extraction evela kwizicubu ziyafuneka, oko kukuthi, i-nucleus ikhutshwa ngokuthe ngqo kwiithishu okanye kwiseli kwaye ilungiswe ibe yi-single-nucleus suspension for single- ulandelelwano lweseli.
I-BMK inikezela nge-10 × i-Genomics yeChromium TM esekelwe kwinkonzo yokulandelelana kwe-RNA yeseli enye.Le nkonzo isetyenziswe ngokubanzi kwizifundo kwizifundo ezinxulumene nesifo, ezifana nokwahlukana kweeseli zomzimba, i-tumor heterogeneity, uphuhliso lwezicubu, njl.
I-Spatial transcriptome chip: 10× Genomics
Iqonga: I-Illumina NovaSeq Platform
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Ulandelelwano lweGenome yeSityalo/yeSilwanyana
Ukulandelelana kwakhona kwe-genome yonke, eyaziwa ngokuba yi-WGS, yenza ukuba kutyhilwe zombini iinguqulelo eziqhelekileyo kunye nezinqabileyo kuyo yonke i-genome kuquka i-Single Nucleotide Polymorphism (SNP), ukucinywa kokuFaka (InDel), ukwahluka koLwakhiwo (SV), kunye neCopy Number Variation (CNV). ).I-SVs zenza inxalenye enkulu yesiseko sokwahluka kune-SNP kwaye ibe nempembelelo enkulu kwi-genome, enempembelelo ebalulekileyo kwizinto eziphilayo.Ukuphinda kufundwe ixesha elide kuvumela ukuchongwa okuchane ngakumbi kwamaqhekeza amakhulu kunye nokwahluka okuntsokothileyo kuba ukufunda ixesha elide kwenza kube lula ukuwela ichromosomal ngaphaya kwemimandla entsonkothileyo efana nokuphindaphinda kwe-tandem, imimandla etyebileyo yeGC/AT, kunye nemimandla eguquguqukayo.
Iqonga: Illumina, PacBio, Nanopore
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I-BMKMANU S1000 i-Spatial Transcriptome
Ukulungelelaniswa kwendawo yeeseli kudlala indima ebalulekileyo kwiinkqubo ezahlukeneyo zebhayoloji, ezifana nokungena komzimba, ukuphuhliswa kwe-embryo, njl.Ngophuhliso lwetekhnoloji, i-ultra-clear-clear tissue morphology kunye nomahluko wokwenyani wesakhiwo sokubonakaliswa kwemolekyuli yendawo kufuneka ifundwe ngesisombululo esiphezulu.I-BMKGENE ibonelela ngenkonzo ebanzi, yokumisa enye yolandelelwano lwe-spatial transcriptome ukusuka kwiisampulu ukuya kwiimbono zebhayoloji.
Itekhnoloji ye-Spatial transcriptomics ixhobisa iimbono zenoveli kwibala lophando olwahlukeneyo ngokusombulula iprofayile yegenesis enomxholo wesithuba kwiisampulu ezingafaniyo.
Itshiphu ye-Spatial transcriptome: BMKMANU S1000
Iqonga: I-Illumina NovaSeq Platform
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I-10x ye-Genomics yeVisium ye-Spatial Transcriptome
I-Visium Spatial Gene Expression yitekhnoloji yolandelelwano lwe-spatial transcriptome yokuhlela izicubu ngokusekelwe kwi-mRNA iyonke.Imephu yonke i-transcriptome kunye nomxholo we-morphological ukufumanisa ulwazi lwenoveli kuphuhliso oluqhelekileyo, izifo zesifo, kunye nophando loguqulo lweklinikhi.I-BMKGENE ibonelela ngenkonzo ebanzi, yokumisa enye yolandelelwano lwe-spatial transcriptome ukusuka kwiisampulu ukuya kwiimbono zebhayoloji.
Itekhnoloji ye-Spatial transcriptomics ixhobisa iimbono zenoveli kwiindawo ezahlukeneyo zophando ngokusombulula iprofayile yegenesis kunye nomxholo wendawo kwiisampulu ezingafaniyo..
I-Spatial transcriptome chip: 10x Genomics Visium
Iqonga:I-Illumina NovaSeq Platform
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Ubude obupheleleyo be-mRNA Ukulandelelana-Nanopore
Ulandelelwano lwe-RNA ibe sisixhobo esixabisekileyo sohlalutyo olupheleleyo lwe-transcriptome.Ngokungathandabuzekiyo, ulandelelwano lokufunda okufutshane lwemveli luzuze uphuhliso oluninzi olubalulekileyo apha.Nangona kunjalo, ihlala idibana nemida ekuchongeni kwe-isoform yobude obugcweleyo, ubungakanani, ukuthambekela kwePCR.
Ulandelelwano lwe-Nanopore luzahlula kwezinye iiplatifomu zokulandelelana, kuba i-nucleotides ifundwa ngokuthe ngqo ngaphandle kwe-DNA synthesis kwaye ivelise ixesha elide lokufunda kwi-kilobases ezimashumi.Oku kuxhobisa ukufundwa ngokuthe ngqo ukuwela imibhalo yobude obugcweleyo kunye nokujongana nemingeni kwizifundo zenqanaba le-isoform.
Iqonga:Nanopore PromethION
Ithala leencwadi:cDNA-PCR
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Ulandelelwano olupheleleyo lwe-mRNA -PacBio
De novoUlandelelwano olupheleleyo lwe-transcriptome, ekwaziwa njengeDe novoI-Iso-Seq ithatha inzuzo ye-PacBio sequencer kubude bokufunda, eyenza ukulandelelana kwee-molecule ze-cDNA ezigcweleyo ngaphandle kokuphazamiseka.Oku kuthintela ngokupheleleyo naziphi na iimpazamo eziveliswe kumanyathelo endibano yokukhutshelwa kwaye kwakha iiseti ze-unigene kunye nesisombululo senqanaba le-isoform.Ezi seti ze-unigene zibonelela ngolwazi olunamandla lwemfuza njenge "reference genome" kwinqanaba le-transcriptome.Ukongeza, ukudibanisa nedatha yolandelelwano lwesizukulwana esilandelayo, le nkonzo ixhobisa ubungakanani obuchanekileyo be-isoform-level expression.
Iqonga: I-PacBio Sequel IIIthala leencwadi: Ilayibrari yentsimbi ye-SMRT -
Eukaryotic mRNA Sequencing-Illumina
Ulandelelwano lwe-mRNA lwenza ukuba iprofayili yazo zonke ii-mRNA zikhutshelwe kwiiseli phantsi kweemeko ezithile.Bubuchwephesha obunamandla bokutyhila inkangeleko yemfuza yemfuza, izakhiwo zemfuza kunye neendlela zemolekyuli zeenkqubo ezithile zebhayoloji.Ukuza kuthi ga ngoku, ulandelelwano lwe-mRNA lusetyenziswe ngokubanzi kuphando olusisiseko, uxilongo lwezonyango, uphuhliso lwamachiza, njl.
Iqonga: I-Illumina NovaSeq Platform
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Non-Reference esekelwe mRNA Sequencing-Illumina
Ulandelelwano lwe-mRNA lusebenzisa ubuchule bolandelelwano lwesizukulwana esilandelayo (NGS) ukubamba umthunywa i-RNA(mRNA) ifom ye-Eukaryote ngexesha elithile lemisebenzi ekhethekileyo eyenziwayo.Owona mbhalo mde uceliweyo wawubizwa ngokuba 'yi-Unigene' kwaye isetyenziswe njengolandelelwano lwereferensi kuhlalutyo olulandelayo, oluyindlela esebenzayo yokufunda umatshini wemolekyuli kunye nothungelwano lolawulo lweentlobo ngaphandle kwereferensi.
Emva kokuhlanganiswa kwedatha ye-transcriptome kunye nochazo olusebenzayo lwe-unigene
(1) Uhlalutyo lwe-SSR, ukubikezelwa kwe-CDS kunye nesakhiwo sofuzo siya kwenziwa kwangaphambili.
(2) Ubungakanani bentetho ye-unigene kwisampulu nganye iya kwenziwa.
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(4) Ukuhlanganisana, uchazo olusebenzayo kunye nohlalutyo lokutyebisa lwee-unigenes ezichazwe ngokwahlukileyo ziya kwenziwa.
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Ixesha elide non-coding ulandelelwano-Illumina
Ii-RNAs ezinde ezingezizo iikhowudi (lncRNAs) luhlobo lweeamolekyuli ze-RNA ezinobude obungaphezulu kwe-200 nt, eziphawulwa ngokwazi ukukhowuda okuphantsi kakhulu.I-LncRNA, njengelungu eliphambili kwii-RNA ezingakhowudi, ifumaneka kakhulu kwi-nucleus kunye ne-plasma.Uphuhliso lolandelelwano lwetekhnoloji kunye ne-bioinformatics yenza ukuba kuchongwe iinoveli ezininzi ze-lncRNAs kwaye zinxulumanise ezo zinemisebenzi yebhayoloji.Ubungqina obuninzi bubonisa ukuba i-lncRNA ibandakanyeka ngokubanzi kulawulo lwe-epigenetic, umgaqo wokubhala kunye nomgaqo wokubhala emva kokubhala.
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I-RNA encinci yokulandelelana-Illumina
I-RNA encinci ibhekisela kwiklasi yee-molecule ze-RNA ezingenayo ikhowudi ezihlala zingaphantsi kwe-200nt ubude, kubandakanywa i-RNA encinci (i-miRNA), i-RNA encinci (i-siRNA), kunye ne-piwi-interacting RNA (piRNA).
I-MicroRNA (miRNA) yiklasi ye-RNA encinci engapheliyo kunye nobude malunga ne-20-24nt, edlala indima ebalulekileyo yokulawula kwiiseli.I-miRNA ibandakanyeka kwiinkqubo ezininzi zobomi ezibonisa izicubu-ezithile kunye nenqanaba - ukubonakaliswa okuthe ngqo kwaye kugcinwe kakhulu kwiintlobo ezahlukeneyo.
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circRNA ulandelelwano-Illumina
I-CircRNAs (i-RNAs yeSetyhula) yiklasi enye yee-molecule ze-RNA ezingenakho ikhowudi ezimfutshane kwi-5′ end cap kunye ne-3′ end poly(A) umsila.Ii-CircRNAs zenza ulwakhiwo lwesetyhula nge-covalent bond ekwimo echasene ne-RNA exonuclease digestion.I-circRNA idlala indima ebalulekileyo ekukhuleni nasekuphuhliseni izinto eziphilayo kunye nokuchasana kwazo kwimeko yangaphandle.
I-CircRNA inemisebenzi emininzi, enokuthi ibophe ngokukhuphisana i-miRNA, isebenzise umsebenzi wokulawula we-ceRNA, ukulawula ukubonakaliswa kofuzo.Kwiminyaka yakutshanje, kuye kwafunyaniswa ukuba inxulumene ngokusondeleyo nesehlo kunye nophuhliso lwezifo, kwaye inamathuba amakhulu okusebenza kwicala leempawu zokuxilonga izifo njalo njalo.
