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I-Genome-wide Association Analysis
Ucwaningo lwe-Genome-wide association (GWAS) luhlose ukuhlonza okuhlukile kofuzo (i-genotype) ehlotshaniswa nezici ezithile (i-phenotype).Ucwaningo lwe-GWAS luphenya izimpawu zofuzo eziwela lonke i-genome yenani elikhulu labantu futhi libikezela izinhlangano ze-genotype-phenotype ngokuhlaziywa kwezibalo ezingeni labantu.Isetshenziswe kabanzi ocwaningweni lwezifo zabantu kanye nokumbiwa kwezakhi zofuzo ezicini eziyinkimbinkimbi zezilwane noma izitshalo.
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I-Single-nucleus RNA Sequencing
Ukuthuthuka ekuthwebuleni iseli elilodwa kanye nezindlela zokwakha umtapo wolwazi ngamunye okuhlanganiswe nokulandelana komphumela ophezulu kuvumela izifundo zesisho sofuzo ngesisekelo seseli-by-cell.Inika amandla ukuhlaziya okujulile nokuphelele kwesistimu kumaseli ayinkimbinkimbi, lapho igwema khona kakhulu ukufihla ukuhlukahluka kwawo ngokuthatha isilinganiso sawo wonke amaseli.
Kodwa-ke, amanye amangqamuzana awafanele ukwenziwa abe yi-single-cell ukumiswa, ngakho-ke ezinye izindlela zokulungiselela isampula - ukukhishwa kwe-nucleus ezicutshini ziyadingeka, okungukuthi, i-nucleus ikhishwa ngokuqondile kumathishu noma ingqamuzana futhi ilungiselwe ukumiswa kwe-nucleus eyodwa ukuze kusetshenziswe eyodwa- ukulandelana kweseli.
I-BMK ihlinzeka ngesevisi yokulandelana kwe-RNA eyi-10× Genomics ChromiumTM.Le nsizakalo isetshenziswe kakhulu ezifundweni eziphathelene nezifo, njengokuhlukaniswa kwamaseli omzimba, i-tumor heterogeneity, ukuthuthukiswa kwezicubu, njll.
I-Spatial transcriptome chip: 10× Genomics
Ipulatifomu: I-Illumina NovaSeq Platform
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Ukulandelana Kohlobo Lonke Lwezitshalo/Izilwane
Ukulandelana kabusha kwe-genome okuphelele, okubuye kwaziwe nge-WGS, kuvumela ukudalulwa kwakho kokubili ukuguqulwa okuvamile nokungavamile kuyo yonke i-genome okuhlanganisa i-Single Nucleotide Polymorphism (SNP), Ukususwa Kokufaka (InDel), Ukuhluka Kwesakhiwo (SV), kanye Nokuhlukahluka Kwenombolo Kopisha (CNV ).Ama-SV enza ingxenye enkulu yesisekelo sokuhlukahluka kune-SNP futhi anomthelela omkhulu ku-genome, enomthelela obalulekile ezintweni eziphilayo.Ukuphinda kufundwe isikhathi eside kuvumela ukuhlonza okunembe kakhudlwana kwezingcezu ezinkulu nokuhluka okuyinkimbinkimbi ngoba ukufunda isikhathi eside kwenza kube lula kakhulu ukweqa i-chromosomal ezindaweni eziyinkimbinkimbi njengokuphindaphinda kwe-tandem, izifunda ezicebile ze-GC/AT, nezifunda eziguquguquka kakhulu.
Ipulatifomu: Illumina, PacBio, Nanopore
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I-BMKMANU S1000 Spatial Transcriptome
Ukuhleleka kwendawo kwamaseli kudlala indima ebalulekile ezinqubweni ezihlukahlukene zebhayoloji, njengokungenwa kwamasosha omzimba, ukuthuthukiswa kombungu, njll. Ukulandelana kwe-Spatial transcriptome, okubonisa iphrofayela yesisho sofuzo kuyilapho kugcinwa ulwazi lwendawo yendawo, kunikeze imininingwane emihle ekwakhiweni kwezicubu zezinga le-transcriptome.Ngokuthuthuka kobuchwepheshe, i-ultra-clear tissue morphology kanye nomehluko wangempela wesakhiwo sokuchazwa kwamangqamuzana wendawo kudingeka kufundwe ngokulungiswa okuphezulu.I-BMKGENE inikeza insizakalo yokulandelana kwe-spatial transcriptome ephelele, yokuma eyodwa ukusuka kumasampula ukuya emininingwaneni yebhayoloji.
Ubuchwepheshe be-Spatial transcriptomics bunike amandla imibono yamanoveli enkundleni yocwaningo eyahlukahlukene ngokuxazulula iphrofayili yesisho sofuzo ngokuqukethwe kwendawo kumasampula ahlukahlukene.
I-Spatial transcriptome chip: BMKMANU S1000
Ipulatifomu: I-Illumina NovaSeq Platform
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I-10x Genomics Visium Spatial Transcriptome
I-Visium Spatial Gene Expression iwubuchwepheshe obujwayelekile bokulandelana kwe-transcriptome yendawo yokuhlukanisa izicubu ngokusekelwe kungqikithi ye-mRNA.Imephu yonke i-transcriptome enomongo we-morphological ukuthola imininingwane yenoveli ekuthuthukisweni okujwayelekile, ukugula kwezifo, kanye nocwaningo lokuhumusha lomtholampilo.I-BMKGENE inikeza insizakalo yokulandelana kwe-spatial transcriptome ephelele, yokuma eyodwa ukusuka kumasampula ukuya emininingwaneni yebhayoloji.
Ubuchwepheshe be-Spatial transcriptomics bunike amandla imibono yamanoveli enkundleni yocwaningo eyahlukene ngokuxazulula iphrofayili yokuvezwa kofuzo ngokuqukethwe kwendawo kumasampuli ahlukahlukene..
I-Spatial transcriptome chip: 10x Genomics Visium
Inkundla:I-Illumina NovaSeq Platform
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Ukulandelana kwe-mRNA yobude obugcwele-Nanopore
Ukulandelana kwe-RNA kube ithuluzi eliyigugu lokuhlaziya okuphelele kwe-transcriptome.Ngokungangabazeki, ukulandelana kwendabuko kokufunda okufushane kuzuze intuthuko eminingi ebalulekile lapha.Noma kunjalo, imvamisa ihlangabezana nemikhawulo ekuhlonzweni kobude obugcwele be-isoform, ukulinganisa, ukuchema kwe-PCR.
Ukulandelana kwe-Nanopore kuzihlukanisa kwezinye izinkundla zokulandelana, ngokuthi ama-nucleotide afundwa ngokuqondile ngaphandle kokuhlanganiswa kwe-DNA futhi akhiqize ukufunda isikhathi eside emashumini ama-kilobase.Lokhu kunika amandla okulotshiweyo okufundwayo okuqondile okuwela ubude obugcwele kanye nokubhekana nezinselele ezifundweni zeleveli ye-isoform.
Inkundla:I-Nanopore PromethION
Umtapowolwazi:cDNA-PCR
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Ukulandelana kwe-mRNA yobude obugcwele -PacBio
De novoukulandelana kwe-transcriptome yobude obugcwele, eyaziwa nangokuthiDe novoI-Iso-Seq ithatha izinzuzo ze-sequencer ye-PacBio ngobude bokufunda, okuvumela ukulandelana kwama-molecule e-cDNA anobude obugcwele ngaphandle kokuphuka.Lokhu kugwema ngokuphelele noma imaphi amaphutha akhiqizwe ezinyathelweni zokuhlanganisa okulotshiweyo futhi kwakha amasethi e-unigene anokulungiswa kwezinga le-isoform.Lawa masethi e-unigene ahlinzeka ngolwazi lofuzo olunamandla njengokuthi "i-reference genome" ezingeni le-transcriptome.Ngaphezu kwalokho, ukuhlanganisa nedatha yokulandelanisa yesizukulwane esilandelayo, le sevisi inika amandla ukulinganisa okunembile kwenkulumo yezinga le-isoform.
Ipulatifomu: I-PacBio Sequel IIUmtapo wolwazi: Umtapo wensimbi we-SMRT -
I-Eukaryotic mRNA Sequencing-Illumina
Ukulandelana kwe-mRNA kunika amandla ukuphrofayela wawo wonke ama-mRNA alotshwe kusuka kumaseli ngaphansi kwezimo ezithile.Ubuchwephesha obunamandla bokuveza iphrofayili yesisho sofuzo, izakhiwo zofuzo nezindlela zamangqamuzana ezinqubo ezithile zebhayoloji.Kuze kube manje, ukulandelana kwe-mRNA sekusetshenziswe kabanzi ocwaningweni oluyisisekelo, ekuxilongweni komtholampilo, ekuthuthukisweni kwezidakamizwa, njll.
Ipulatifomu: I-Illumina NovaSeq Platform
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I-Non-Reference based mRNA Sequencing-Illumina
Ukulandelana kwe-mRNA kusebenzisa inqubo yesizukulwane esilandelayo sokulandelanisa (NGS) ukuze ithwebule i-RNA(mRNA) yesithunywa ifomu i-Eukaryote ngesikhathi esithile imisebenzi ethile ekhethekile eyenza isebenze.Umbhalo omude kakhulu ohlukanisiwe wawubizwa ngokuthi 'i-Unigene' futhi wasetshenziswa njengochungechunge lwereferensi lokuhlaziya okwalandela, okuyindlela ephumelelayo yokufunda indlela yamangqamuzana nenethiwekhi yokulawula yohlobo ngaphandle kwereferensi.
Ngemuva kokuhlanganiswa kwedatha ye-transcriptome kanye nesichasiselo esisebenzayo se-unigene
(1) Ukuhlaziywa kwe-SSR, ukubikezela kwe-CDS kanye nesakhiwo sofuzo kuzokwenziwa kusengaphambili.
(2) Ukulinganisa kwenkulumo ye-unigene kusampula ngayinye kuzokwenziwa.
(3) Ama-unigene avezwe ngokuhlukile phakathi kwamasampula (noma amaqembu) azotholwa ngokusekelwe ekukhulumeni okungafani
(4) Ukuhlanganisa, isichasiselo esisebenzayo kanye nokuhlaziywa kokunothisa kwama-unigene avezwe ngokwehlukana kuzokwenziwa.
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Ukulandelana okungafaki amakhodi okude-Illumina
Ama-RNA amade angafaki amakhodi (lncRNAs) awuhlobo lwama-molecule e-RNA anobude obudlula ama-200 nt, abonakala ngamandla aphansi kakhulu okubhala amakhodi.I-LncRNA, njengelungu eliyinhloko kuma-RNA angafaki amakhodi, itholakala kakhulu ku-nucleus ne-plasma.Ukuthuthukiswa kobuchwepheshe bokulandelana kanye ne-bioinformatics kwenza ukukhonjwa kwamanoveli amaningi e-lncRNA futhi kuhlobanise lawo anemisebenzi yebhayoloji.Ubufakazi obunqwabelene buphakamisa ukuthi i-lncRNA ihileleke kakhulu ekulawuleni i-epigenetic, ukulawulwa kokulotshiweyo kanye nomthethonqubo wangemva kokubhala.
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Ukulandelana kwe-RNA encane-Illumina
I-RNA encane ibhekisela ekilasini lama-molecule e-RNA angafaki amakhodi ngokuvamile angaphansi kuka-200nt ngobude, okuhlanganisa i-RNA encane (miRNA), ukuphazamiseka okuncane kwe-RNA (siRNA), kanye ne-piwi-interacting RNA (piRNA).
I-MicroRNA (miRNA) ikilasi le-RNA encane engokwemvelo enobude obungaba ngu-20-24nt, edlala izindima ezibalulekile zokulawula kumaseli.I-miRNA ibandakanyeka ezinqubweni eziningi zokuphila eziveza izicubu - eziqondile kanye nesiteji - inkulumo ethize futhi egcinwe kakhulu ezinhlotsheni ezahlukene.
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i-circRNA sequencing-Illumina
I-CircRNAs (Circular RNAs) yisigaba esisodwa sama-molecule e-RNA angafaki amakhodi afushane ngo-5′ end cap kanye no-3′ end poly(A) umsila.Ama-CircRNA enza isakhiwo esiyindilinga ngesibopho esihlangene okuyisimo esimelene nokugaya kwe-RNA exonuclease.I-circRNA idlala indima ebalulekile ekukhuleni nasekuthuthukisweni kwezinto eziphilayo kanye nokumelana kwazo nemvelo yangaphandle.
I-CircRNA inemisebenzi eminingi, engahlanganisa i-miRNA ngokuncintisana, isebenzise umsebenzi wokulawula we-ceRNA, ukuze ilawule ukubonakaliswa kofuzo.Eminyakeni yamuva, kuphinde kwatholakala ukuthi kuhlobene eduze nokuvela nokuthuthukiswa kwezifo, futhi kunamathuba amahle okufaka isicelo ekuqondeni izimpawu zokuxilongwa kwezifo nokunye.
