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I-BMKMANU S3000_Spatial Transcriptome
I-Spatial transcriptomics yindlela esivumela ukuba sibambe kwaye sibone ukubonakaliswa kwezakhi zofuzo ngaphakathi kwizicubu. Oku kunokuba luncedo kakhulu ukuqonda indlela iiseli ezisebenzisana ngayo.
Kukho amaqonga ahlukeneyo ale ndlela. Kulo mba, i-BMKGene iphuhlise i-BMKManu 3000 Spatial transcriptome Chip, iqonga eliphucula ukusebenza kobugcisa, lifikelela kwisisombululo esingaphantsi kweeseli kwaye livumela useto lwesisombululo esinamanqanaba amaninzi.
Le chip ifaka amabala ayi-4.2 yezigidi isebenzisa ubuchwepheshe obunelungelo lobunikazi bee-microwells ezifakwe ii-beads ezigcwele ii-probes ezine-barcode. Ngale ndlela, emva kokubanjwa nokwandiswa, sifumana ithala leencwadi le-cDNA elinongiweyo ngeesampulu ezine-barcode ezihambelana ne-Illumina.
Kwidatha, indibaniselwano yebarcode yendawo kunye nee-UMIs iqinisekisa ukuchaneka kunye nokuchaneka kwedatha evelisiweyo. Ukudibanisa konke okungentla, i-BMKManu ibonelela ngoseto lwedatha oluguquguqukayo kakhulu.
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Ulandelelwano lwe-RNA ye-nuclei enye
Uphuhliso lweendlela zokubamba iiseli enye kunye nokwakha ilayibrari ngokwezifiso, kunye nokulandelelana okuphezulu, kutshintshe izifundo zokubonakaliswa kweejini kwinqanaba leseli. Olu phuculo luvumela uhlalutyo olunzulu nolupheleleyo lwamaqela eeseli antsonkothileyo, ukoyisa imida enxulumene nokubonakaliswa kwejini okuqhelekileyo kuzo zonke iiseli kunye nokugcina ukungafani kokwenyani kwezi qela. Ngelixa ulandelelwano lwe-RNA yeseli enye (scRNA-seq) luneengenelo ezingenakuphikiswa, luhlangabezana nemingeni kwiithishu ezithile apho ukudalwa kokumiswa kweseli enye kubonakala kunzima kwaye kufuna iisampuli ezintsha. Kwi-BMKGene, sijongana nalo mqobo ngokunikezela ngokulandelana kwe-RNA ye-nucleus enye (snRNA-seq) sisebenzisa itekhnoloji ye-10X Genomics Chromium yanamhlanje. Le ndlela yandisa uluhlu lweesampuli ezilungele uhlalutyo lwe-transcriptome kwinqanaba leseli enye.
Ukuhlukaniswa kweenuclei kwenziwa nge-chip entsha ye-10X Genomics Chromium, enenkqubo ye-microfluidics yeendlela ezisibhozo ene-double crossings. Ngaphakathi kwale nkqubo, ii-gel beads eziquka ii-barcodes, ii-primers, ii-enzymes, kunye ne-nucleus enye zifakwe kwii-oil drops ezinobukhulu be-nanoliter, zenze i-Gel Bead-in-Emulsion (GEM). Emva kokwakhiwa kwe-GEM, i-cell lysis kunye nokukhululwa kwe-barcode kwenzeka ngaphakathi kwe-GEM nganye. Emva koko, ii-mRNA molecules ziguqulelwa kwi-reverse transcription zibe yi-cDNAs, zifaka ii-barcodes ze-10X kunye ne-Unique Molecular Identifiers (UMIs). Ezi cDNAs emva koko ziphantsi kolwakhiwo lwelayibrari oluqhelekileyo, oluququzelela uphando oluqinileyo nolupheleleyo lweeprofayili zokubonakaliswa kwezakhi zofuzo kwinqanaba leseli enye.
Iqonga: Iqonga le-10 × Genomics Chromium kunye ne-Illumina NovaSeq
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I-10x Genomics Visium Spatial Transcriptome
I-Spatial transcriptomics yiteknoloji entsha evumela abaphandi ukuba baphande iipateni zokubonakaliswa kwezakhi zofuzo ngaphakathi kwezicubu ngelixa begcina imeko yazo yendawo. Elinye iqonga elinamandla kweli candelo yi-10x Genomics Visium edityaniswe nokulandelelana kwe-Illumina. Umgaqo we-10X Visium usekwe kwi-chip ekhethekileyo enendawo yokubamba ekhethiweyo apho kubekwa khona amacandelo ezicubu. Le ndawo yokubamba iqulethe amabala aneebhakhowudi, ngalinye lihambelana nendawo eyahlukileyo ngaphakathi kwezicubu. Iimolekyuli ze-RNA ezibanjiweyo ezivela kwizicubu emva koko ziphawulwa ngee-unique molecular identifiers (UMIs) ngexesha lenkqubo yokubhala engasemva. La mabala aneebhakhowudi kunye nee-UMI zivumela ukumapha ngokuchanekileyo kwendawo kunye nokulinganisa ukubonakaliswa kwezakhi zofuzo kwisisombululo seseli enye. Ukudibanisa iisampuli ezineebhakhowudi kunye nee-UMI kuqinisekisa ukuchaneka kunye nokuchaneka kwedatha evelisiweyo. Ngokusebenzisa obu buchwepheshe be-Spatial Transcriptomics, abaphandi banokufumana ukuqonda okunzulu malunga nokuhlelwa kwendawo kweeseli kunye nokusebenzisana okuntsonkothileyo kweemolekyuli okwenzeka ngaphakathi kwezicubu, okubonelela ngengqiqo ebalulekileyo kwiinkqubo ezisisiseko seenkqubo zebhayoloji kwiindawo ezininzi, kubandakanya i-oncology, i-neuroscience, i-biology yophuhliso, i-immunology, kunye nezifundo zezityalo.
Iqonga: 10X Genomics Visium kunye ne-Illumina NovaSeq
