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I-Chromatin Immunoprecipitation Sequencing (ChIP-seq)
I-ChIP-Seq inikeza iphrofayili ebanzi yofuzo lwezinjongo ze-DNA zokuguqulwa kwe-histone, izici zokuloba, namanye amaprotheni ahlobene ne-DNA.Ihlanganisa ukukhethwa kwe-chromatin immuno-precipitation (ChIP) ukuze kubuyiselwe ama-protein-DNA complexes athile, namandla okulandelana kwesizukulwane esilandelayo (NGS) okulandelana okuphezulu komphumela we-DNA etholiwe.Ukwengeza, ngenxa yokuthi ama-protein-DNA complexes atholakala kumangqamuzana aphilayo, izingosi ezibophayo zingaqhathaniswa ngezinhlobo zamangqamuzana nezicubu ezihlukene, noma ngaphansi kwezimo ezihlukene.Izicelo zisukela ekulawuleni okulotshiweyo kuye ezindleleni zokuthuthuka eziya ezindleleni zezifo nangale kwalokho.
Ipulatifomu: I-Illumina NovaSeq Platform
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Ukulandelana kwe-Metagenomic -NGS
I-Metagenome ibhekisela eqoqweni lengqikithi yezinto zofuzo zomphakathi oxubile wezinto eziphilayo, njenge-metagenome yemvelo, i-metagenome yomuntu, njll. Iqukethe ama-genome akho kokubili amagciwane aphilayo nangatshalwa.Ukulandelana kwe-Metagenomic iyithuluzi lamangqamuzana elisetshenziselwa ukuhlaziya izinto ezixubile ze-genomic ezikhishwe kumasampula emvelo, okunikeza ulwazi oluningiliziwe ngokuhlukahluka kwezinhlobo kanye nobuningi, ukwakheka kwabantu, ubudlelwano be-phylogenetic, izakhi zofuzo ezisebenzayo kanye nenethiwekhi yokuxhumana nezici zemvelo.
Inkundla:I-Illumina NovaSeq Platform
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Ukulandelana kwe-Metagenomic-Nanopore
I-Metagenomics iyithuluzi lamangqamuzana elisetshenziselwa ukuhlaziya izinto ezixubile ze-genomic ezikhishwe kumasampula emvelo, ezihlinzeka ngolwazi oluningiliziwe ngokuhlukahluka kwezinhlobo kanye nobuningi, ukwakheka kwabantu, ubudlelwano be-phylogenetic, izakhi zofuzo ezisebenzayo kanye nenethiwekhi yokuxhumana enezici zemvelo, njll. Amapulatifomu okulandelana kwe-Nanopore asanda kwethulwa kwizifundo ze-metagenomic.Ukusebenza kwayo okuhle kakhulu kubude bokufunda kuthuthukise kakhulu ukuhlaziywa kwe-metagenomic yokusakaza, ikakhulukazi ukuhlanganiswa kwe-metagenome.Ngokuthatha izinzuzo zobude bokufunda, ucwaningo lwe-metagenomic olusekelwe ku-Nanopore luyakwazi ukuzuza umhlangano oqhubekayo uma uqhathaniswa ne-shot-gun metagenomics.Kushicilelwe ukuthi i-metagenomics esekwe e-Nanopore ikhiqize ngempumelelo ama-genomes ebhaktheriya aphelele futhi avaliwe avela kuma-microbiomes (Moss, EL, et. al,I-Nature Biotech, 2020)
Inkundla:I-Nanopore PromethION P48
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Ukulandelana okuphelele kwe-genome bisulfite
I-DNA methylation endaweni yesihlanu ku-cytosine (5-mC) inethonya elibalulekile ekukhulumeni kwezakhi zofuzo kanye nomsebenzi wamaselula.Amaphethini e-methylation angavamile ahlotshaniswa nezimo nezifo eziningana, njengomdlavuza.I-WGBS isiphenduke indinganiso yegolide yokufunda i-genome-wide methylation ngokulungiswa kwesisekelo esisodwa.
Ipulatifomu: I-Illumina NovaSeq Platform
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I-Assay ye-Chromatin Efinyelelekayo Ye-Transposase Enokulandelanisa Okuphezulu Kokudlula (i-ATAC-seq)
I-ATAC-seq iyindlela yokulandelanisa ephezulu yokuhlaziya ukufinyeleleka kwe-chromatin yohlobo lonke, okubalulekile ekulawuleni i-epigenetic yomhlaba wonke ekukhulumeni kofuzo.Ama-adaptha alandelanayo afakwa ezifundeni ze-chromatin nge-hyperactive Tn5 transposase.Ngemuva kokukhulisa i-PCR, kwakhiwa umtapo wolwazi wokulandelanisa.Zonke izifunda ze-chromatin ezivulekile zingatholwa ngaphansi kwesimo esithile sesikhathi sesikhala, hhayi kuphela kumasayithi abophezelayo we-transcription factor, noma isifunda esithile esilungisiwe se-histone.
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16S/18S/ITS Amplicon Sequencing-PacBio
I-subunit ku-16S kanye ne-18S rRNA equkethe zombili izifunda ezilondolozwe kakhulu neziguquguqukayo iwuphawu lweminwe oluphelele lwemolekyuli yokuhlonza i-prokaryotic kanye ne-eukaryotic organisms.Ngokusebenzisa ithuba lokulandelanisa, lawa ma-amplicon angaqondiswa ngokusekelwe ezingxenyeni ezilondoloziwe futhi izifunda eziguquguqukayo zingabonakaliswa ngokugcwele ukuhlonza i-microbial enomthelela ocwaningweni oluhlanganisa ukuhlaziywa kwe-microbial diversity, taxonomy, phylogeny, njll. I-molecule ye-Single-real-time(SMRT) ) ukulandelana kweplathifomu ye-PacBio kwenza kutholwe ukufundwa okude okunembe kakhulu, okungahlanganisa ama-amplicon anobude obugcwele (cishe u-1.5 KB).Ukubuka okubanzi kwendima yofuzo kuthuthukise kakhulu ukulungiswa kwesichasiselo sezinhlobo kubhaktheriya noma emphakathini wesikhunta.
Inkundla:I-PacBio Sequel II
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16S/18S/ITS Amplicon Sequencing-NGS
Ukulandelana kwe-amplicon ye-16S/18S/ITS kuhloswe ekudaluleni i-phylogeny, taxonomy, kanye nezinhlobo eziningi emphakathini we-microbial ngokuphenya imikhiqizo ye-PCR yezimpawu zofuzo zokugcina izindlu eziqukethe zombili izingxenye ezixoxwa kakhulu nezingaguquguquki.Ukwethulwa kwalezi zigxivizo zeminwe zamangqamuzana aphelele ngu-Woeses et al,(1977) kunika amandla ukuhlukaniswa kwephrofayili ye-microbiome.Ukulandelana kwe-16S (ibhaktheriya), i-18S (isikhunta) kanye ne-Internal transcribed spacer(ITS, isikhunta) kuvumela ukukhonjwa kwazo zombili izinhlobo zezinhlobo eziningi kanye nezinhlobo ezingavamile nezingaziwa.Lobu buchwepheshe sebuphenduke ithuluzi elisetshenziswa kabanzi futhi elikhulu ekuhlonzeni ukwakheka kwe-microbial ehlukile ezindaweni ezahlukahlukene, njengomlomo womuntu, amathumbu, indle, njll.
Inkundla:I-Illumina NovaSeq Platform
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Ukulandelana Kabusha Kwe-Genome Yamagciwane kanye Nesikhunta
Ukulandelana kabusha kwe-genome yegciwane kanye nesikhunta kuyithuluzi elibalulekile lokuqedela ama-genomes amagciwane aziwayo nesikhunta, kanye nokuqhathanisa ama-genome amaningi noma ukwenza imephu yezakhi zezinto eziphilayo ezintsha.Kubaluleke kakhulu ukulandelana kwawo wonke ama-genome e-bacterium nesikhunta ukuze kukhiqizwe ama-genome ayinkomba anembile, ukwenza ukuhlonza ama-microbial kanye nezinye izifundo zokuqhathanisa zofuzo.
I-Platform:I-Illumina NovaSeq Platform
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I-PacBio-Full-length 16S/18S/ITS Amplicon Sequencing
Iplathifomu ye-Amplicon (16S/18S/ITS) yathuthukiswa yaba neminyaka yokuhlangenwe nakho ekuhlaziyweni kwephrojekthi ye-microbial diversity, equkethe ukuhlaziya okuyisisekelo okujwayelekile kanye nokuhlaziya komuntu siqu: ukuhlaziya okuyisisekelo kuhlanganisa okuqukethwe kokuhlaziya okujwayelekile kocwaningo lwamanje lwe-microbial, okuqukethwe kokuhlaziya kucebile futhi kubanzi, futhi imiphumela yokuhlaziya yethulwa ngendlela yemibiko yephrojekthi;Okuqukethwe kokuhlaziywa komuntu siqu kuhlukahlukene.Amasampuli angakhethwa futhi amapharamitha angasethwa ngendlela evumelana nezimo ngokuvumelana nombiko wokuhlaziya oyisisekelo kanye nenjongo yocwaningo, ukuze kufezeke izidingo eziqondene nawe.Isistimu yokusebenza yeWindows, elula futhi esheshayo.
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I-PacBio-Full-length Transcriptome (Non-Reference)
Ithatha idatha yokulandelanisa ye-Pacific Biosciences (PacBio) Isoform njengokufakwayo, lolu hlelo lokusebenza luyakwazi ukubona ukulandelana kokulotshiweyo kobude obugcwele (ngaphandle kokuhlanganisa).Ngokwenza imephu ukulandelana kobude obugcwele ngokumelene ne-reference genome, okulotshiweyo kungenziwa ngcono ngezakhi zofuzo ezaziwayo, imibhalo, izifunda zekhodi, njll. Kulokhu, ukuhlonza okunembe kakhudlwana kwezakhiwo ze-mRNA, njengokunye ukuhlanganisa, njll, kungafinyelelwa.Ukuhlaziywa okuhlangene nedatha yokulandelanisa ye-NGS transcriptome kunika amandla isichasiselo esibanzi kanye nokulinganisa okunembe kakhudlwana ekukhulumeni ezingeni lokuloba, okuzuzisa kakhulu ukuvezwa komehluko ongezansi nokuhlaziywa kokusebenza.
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I-Reduced Representation Bisulfite Sequencing (RRBS)
Ucwaningo lwe-DNA methylation belulokhu luyisihloko esishisayo ocwaningweni lwezifo, futhi luhlobene kakhulu nezakhi zofuzo kanye nezici ze-pheno-typic.I-RRBS iyindlela enembile, ephumelelayo neyongayo yocwaningo lwe-DNA methylation.Ukunothiswa kwezifunda eziqhingini zomgqugquzeli kanye ne-CpG nge-enzymatic cleavage (Msp I), kuhlanganiswe nokulandelana kwe-Bisulfite, kunikeza ukutholwa kwe-DNA methylation yokuxazulula okuphezulu.
Ipulatifomu: I-Illumina NovaSeq Platform
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Ukulandelana kwe-Prokaryotic mRNA
Ukulandelana kwe-mRNA kunika amandla ukuphrofayili okuphelele kwayo yonke imibhalo ye-mRNA ngaphakathi kwamaseli ngaphansi kwezimo ezithile.Lobu buchwepheshe obusezingeni eliphezulu busebenza njengethuluzi elinamandla, lembula amaphrofayli ayinkimbinkimbi enkulumo yofuzo, izakhiwo zofuzo, nezindlela zamangqamuzana ezihambisana nezinqubo ezihlukahlukene zebhayoloji.Yamukelwe kabanzi ocwaningweni oluyisisekelo, ekuxilongweni komtholampilo, nasekuthuthukisweni kwezidakamizwa, ukulandelana kwe-mRNA kunikeza imininingwane ebunkingeni bokuguquguquka kwamaselula kanye nokulawulwa kofuzo.Ukucubungula kwethu kwesampula ye-prokaryotic mRNA kwenzelwe imibhalo ye-prokaryotic, ebandakanya ukuncipha kwe-rRNA kanye nokulungiselela umtapo wolwazi.
Ipulatifomu: Illumina NovaSeq X
