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I-BMKMANU S3000_Spatial Transcriptome
I-Spatial transcriptomics iyindlela esivumela ukuthi sibambe futhi sibone ngeso lengqondo ukubonakaliswa kwezakhi zofuzo ngaphakathi kwezicubu. Lokhu kungabaluleka kakhulu ukuqonda ukuthi amaseli asebenzisana kanjani.
Kunezinkundla ezahlukene zale ndlela. Kulokhu, i-BMKGene ithuthukise i-BMKManu 3000 Spatial transcriptome Chip, inkundla ethuthukisa ukusebenza kobuchwepheshe, efinyelela isixazululo esingaphansi kwamangqamuzana futhi evumela ukulungiselelwa kwesisombululo esinamazinga amaningi.
Le chip ifaka izindawo eziyizigidi ezingu-4.2 isebenzisa ubuchwepheshe obunelungelo lobunikazi bama-microwell ahlanganiswe nobuhlalu obugcwele ama-probe anebhakhodi yendawo. Ngale ndlela, ngemva kokubanjwa nokwandiswa, sithola umtapo wezincwadi we-cDNA ocebile ngamasampula anebhakhodi ahambisana ne-Illumina.
Kudatha, inhlanganisela yebhakhodi yendawo kanye nama-UMI iqinisekisa ukunemba nokucaciswa kwedatha ekhiqizwe. Uma ihlanganisa konke okungenhla, i-BMKManu inikeza isethingi yedatha eguquguqukayo kakhulu.
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Ukulandelana kwe-RNA ye-nuclei eyodwa
Ukuthuthukiswa kobuchwepheshe bokubamba iseli elilodwa kanye nokwakha umtapo wolwazi ngokwezifiso, kuhlanganiswe nokulandelana okuphezulu, kushintshe izifundo zokubonakaliswa kwezakhi zofuzo ezingeni leseli. Lokhu kuphumelela kuvumela ukuhlaziywa okujulile nokuphelele kwenani lamaseli ayinkimbinkimbi, ukunqoba imikhawulo ehlotshaniswa nokulinganiswa kokubonakaliswa kwezakhi zofuzo kuwo wonke amaseli nokugcina ukungafani kwangempela ngaphakathi kwala maqembu. Ngenkathi ukulandelana kwe-RNA yeseli elilodwa (scRNA-seq) kunezinzuzo ezingenakuphikwa, kuhlangana nezinselele ezicutshini ezithile lapho ukudalwa kokumiswa kweseli elilodwa kuba nzima futhi kudinga amasampula amasha. Kwa-BMKGene, sibhekana nalesi sithiyo ngokunikeza ukulandelana kwe-RNA ye-nucleus eyodwa (snRNA-seq) sisebenzisa ubuchwepheshe be-10X Genomics Chromium obusezingeni eliphezulu. Le ndlela yandisa ububanzi bamasampula afanelekela ukuhlaziywa kwe-transcriptome ezingeni leseli elilodwa.
Ukuhlukaniswa kwama-nuclei kufezwa nge-chip entsha ye-10X Genomics Chromium, equkethe uhlelo lwe-microfluidics oluneziteshi eziyisishiyagalombili oluneziphambano ezimbili. Ngaphakathi kwalolu hlelo, ama-gel beads afaka ama-barcode, ama-primer, ama-enzyme, kanye ne-nucleus eyodwa afakwe emaconsini kawoyela anobukhulu be-nanoliter, akha i-Gel Bead-in-Emulsion (GEM). Ngemva kokwakheka kwe-GEM, i-cell lysis kanye nokukhululwa kwe-barcode kwenzeka ngaphakathi kwe-GEM ngayinye. Ngemva kwalokho, ama-molecule e-mRNA adlula ekubhalweni okuphambene abe ama-cDNA, afaka ama-barcode ayi-10X kanye nama-Unique Molecular Identifiers (UMIs). Lawa ma-cDNA abe esefakwa ekwakhiweni komtapo wolwazi ojwayelekile, okwenza kube lula ukuhlola okuqinile nokuphelele kwamaphrofayili okubonakaliswa kwezakhi zofuzo ezingeni leseli elilodwa.
Ipulatifomu: Ipulatifomu ye-10 × Genomics Chromium kanye ne-Illumina NovaSeq
