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BMKMANU S3000_Spatial Transcriptome
I-Spatial transcriptomics imi phambili kubuchule obutsha bezenzululwazi, ixhobisa abaphandi ukuba baphengulule kwiipateni ezintsonkothileyo zokubonakaliswa kofuzo ngaphakathi kwezicubu ngelixa begcina imeko yabo yendawo. Phakathi kwamaqonga ahlukeneyo, i-BMKGene iphuhlise i-BMKManu S3000 ye-Spatial Transcriptome Chip, iqhayisa ngesisombululo esiphuculweyo se-3.5µm, ifikelela kuluhlu oluncinci, kwaye yenza ukuba useto lwesisombululo senqanaba elininzi. Itshiphu ye-S3000, enamabala amalunga nezigidi ezi-4, isebenzisa i-microwells elelekwe ngamaso alayishwe nge-spatially barcoded capture probes. Ilayibrari ye-cDNA, etyetyiswe ngeebhakhowudi zendawo, ilungiswa ukusuka kwitshiphu ye-S3000 kwaye emva koko ilandelelaniswe kwiqonga le-Illumina NovaSeq. Ukudityaniswa kweesampulu ezinebhakhowudi yendawo kunye nee-UMIs ziqinisekisa ukuchaneka kunye nokuchaneka kwedatha eyenziweyo. I-chip ye-BMKManu S3000 iguquguquka kakhulu, ibonelela ngesisombululo semigangatho emininzi esinokulungiswa kakuhle kwiithishu ezahlukeneyo kunye namanqanaba afunekayo eenkcukacha. Oku kuguquguquka kubeka itshiphu njengokhetho olubalaseleyo lwezifundo ezahlukeneyo zoshicilelo lwesithuba, ukuqinisekisa ukudityaniswa kwendawo echanekileyo kunye nengxolo encinci. Ukusetyenziswa kwetekhnoloji yokwahlulahlula iiseli kunye ne-BMKManu S3000 yenza ukucandwa kwedatha yokubhala kwimida yeeseli, okukhokelela kuhlalutyo olunentsingiselo yebhayoloji ngqo. Ngaphezu koko, isisombululo esiphuculweyo se-S3000 siphumela kwinani eliphezulu leemfuza kunye ne-UMIs echongiwe ngeseli nganye, eyenza uhlalutyo oluchanekileyo kakhulu lweepateni zokushicilela kwendawo kunye nokuhlanganiswa kweeseli.
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Single- nuclei RNA Ulandelelwano
Uphuhliso lokubanjwa kweseli enye kunye nobuchule bokwakha ithala leencwadi, ludityaniswa nolandelelwano oluphezulu, luguqule izifundo zokuchazwa kofuzo kwinqanaba leseli. Le mpumelelo ivumela uhlalutyo olunzulu nolubanzi ngakumbi lweeseli ezintsonkothileyo, ukoyisa imida eyayanyaniswa ne-avareji yembonakalo yemfuza kuzo zonke iiseli kunye nokugcina iheterogeneity yokwenyani phakathi kwaba bantu. Ngelixa ulandelelwano lwe-RNA yeseli enye (scRNA-seq) ineengenelo ezingenakuphikiswa, ihlangabezana nemiceli mngeni kwizicubu ezithile apho ukudalwa kokumiswa kweseli enye kubonakala kunzima kwaye kufuna iisampuli ezintsha. Kwi-BMKGene, sijongana nalo mqobo ngokunikezela ngolandelelwano lwe-nucleus ye-RNA enye (snRNA-seq) sisebenzisa itekhnoloji ye-10X Genomics Chromium yanamhlanje. Le ndlela yokwandisa i-spectrum yeesampuli ezivumelekileyo kuhlalutyo lwe-transcriptome kwinqanaba leseli enye.
Ukwahlukaniswa kwe-nuclei kufezekiswa nge-10X ye-Genomics ye-Chromium chip entsha, equkethe inkqubo ye-microfluidics yamajelo asibhozo kunye nokuwela kabini. Ngaphakathi kwesi sixokelelwano, ubuhlalu be-gel obubandakanya i-barcodes, i-primers, i-enzymes, kunye ne-nucleus enye ifakwe kwi-nanoliter-sized oil drops, eyenza i-Gel Bead-in-Emulsion (GEM). Ukulandela ukubunjwa kwe-GEM, i-cell lysis kunye nokukhululwa kwebhakhowudi kwenzeka ngaphakathi kwe-GEM nganye. Emva koko, iimolekyuli ze-mRNA ziphinda zikhutshelwe umva kwii-cDNAs, ezibandakanya i-10X yeebhakhowudi kunye ne-Unique Molecular Identifiers (UMIs). Ezi cDNAs ke ngoko ziphantsi kolwakhiwo lwethala leencwadi lolandelelwano olusemgangathweni, luququzelela ukuphononongwa okuqinileyo nolubanzi lweeprofayili zokuchazwa kofuzo kwinqanaba leseli enye.
Iqonga: 10× Genomics Chromium kunye Illumina NovaSeq Platform
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I-10x ye-Genomics yeVisium ye-Spatial Transcriptome
I-Spatial transcriptomics yitekhnoloji enqamlezileyo evumela abaphandi ukuba baphande iipatheni zokubonakaliswa kofuzo ngaphakathi kwezicubu ngelixa begcina imeko yabo yendawo. Iqonga elinye elinamandla kule domain yi-10x Genomics Visium edityaniswa nokulandelelana kwe-Illumina. Umgaqo we-10X Visium ulele kwi-chip ekhethekileyo enendawo ekhethiweyo yokubamba apho amacandelo e-tissue abekwe khona. Le ndawo yokubamba iqulethe iindawo ezinebhakhowudi, nganye ihambelana nendawo eyodwa ngaphakathi kwethishu. Iimolekyuli ze-RNA ezibanjiweyo kwiithishu zibhalwa emva koko zibhalwe nge-molecular identifiers (UMIs) ezikhethekileyo ngexesha lenkqubo yokubhala umva. Ezi ndawo zinebhakhowudi kunye nee-UMIs zenza imephu yendawo echanekileyo kunye nobungakanani bokubonakaliswa kofuzo kwisisombululo seseli enye. Ukudityaniswa kweesampulu ezinebhakhowudi yendawo kunye nee-UMIs ziqinisekisa ukuchaneka kunye nokuchaneka kwedatha eyenziweyo. Ngokusebenzisa le teknoloji ye-Spatial Transcriptomics, abaphandi banokufumana ukuqonda okunzulu malunga nentlangano yendawo yeeseli kunye nentsebenziswano eyinkimbinkimbi ye-molecular eyenzeka ngaphakathi kwezicubu, inika ulwazi oluxabisekileyo kwiindlela eziphantsi kweenkqubo zebhayoloji kwiinkalo ezininzi, kuquka i-oncology, neuroscience, i-biology yophuhliso, i-immunology. , kunye nezifundo zebhotani.
Iqonga: I-10X Genomics Visium kunye ne-Illumina NovaSeq
