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I-BMKMANU S3000_Spatial Transcriptome
I-Spatial transcriptomics imi phambili ekusungulweni kwesayensi, inika amandla abacwaningi ukuthi bahlole amaphethini enkulumo eyinkimbinkimbi yezakhi zofuzo phakathi kwezicubu kuyilapho belondoloza isimo sabo sendawo. Phakathi kwamapulatifomu ahlukahlukene, i-BMKGene ithuthukise i-BMKManu S3000 Spatial Transcriptome Chip, eziqhayisa ngokulungiswa okuthuthukisiwe kwe-3.5µm, efinyelela ububanzi obuncane, futhi ivumela izilungiselelo zokuxazulula amazinga amaningi. I-chip ye-S3000, equkethe cishe izindawo eziyizigidi ezi-4, isebenzisa ama-microwell agqitshwe ngobuhlalu obulayishwe ama-probe wokuthwebula anebhakhodi. Umtapo wezincwadi we-cDNA, ohlanganiswe namabhakhodi wendawo, ulungiswa kusukela ku-chip ye-S3000 bese ulandelana ngokulandelana endaweni ye-Illumina NovaSeq. Inhlanganisela yamasampula anebhakhodi yendawo kanye nama-UMI kuqinisekisa ukunemba nokucaciswa kwedatha ekhiqizwayo. I-chip ye-BMKManu S3000 iguquguquka kakhulu, inikezela ngezilungiselelo zokuxazulula amazinga amaningi ezingashunwa kahle kumathishu ahlukene kanye namazinga afiswayo emininingwane. Lokhu kuzivumelanisa nezimo kubeka i-chip njengenketho evelele yezifundo ezihlukahlukene ze-spatial transcriptomics, iqinisekisa ukuhlangana okunembayo kwendawo okunomsindo omncane. Ukusetshenziswa kobuchwepheshe bokuhlukanisa amaseli nge-BMKManu S3000 kuvumela ukunqunywa kwedatha yokubhala emikhawulweni yamaseli, okuholela ekuhlaziyweni okunencazelo eqondile yebhayoloji. Ngaphezu kwalokho, ukulungiswa okuthuthukisiwe kwe-S3000 kuphumela enanini eliphezulu lezakhi zofuzo kanye nama-UMI atholiwe ngeseli ngalinye, okuvumela ukuhlaziya okunembe kakhulu kwamaphethini okulotshwa kwendawo nokuhlanganisa amaseli.
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I-Single-nuclei RNA Sequencing
Ukuthuthukiswa kokuthwebula iseli elilodwa nezindlela zokwakha umtapo wolwazi wangokwezifiso, kuhlanganiswe nokulandelana komphumela ophezulu, kuguqule izifundo zesisho sofuzo ezingeni leseli. Lokhu kuphumelela kuvumela ukuhlaziywa okujulile nokubanzi kakhulu kwezibalo zamangqamuzana ayinkimbinkimbi, kweqa imikhawulo ehambisana nokuvezwa kwezakhi zofuzo okumaphakathi kuwo wonke amaseli kanye nokulondoloza ukuhlukahluka kwangempela phakathi kwalezi zibalo. Nakuba ukulandelana kwe-RNA yeseli elilodwa (scRNA-seq) kunezinzuzo ezingenakuphikwa, kuhlangabezana nezinselele kumathishu athile lapho ukwakhiwa kokumiswa kweseli elilodwa kuba nzima futhi kudinga amasampula amasha. Kwa-BMKGene, sibhekana nalesi sithiyo ngokunikeza ukulandelana kwe-nucleus eyodwa ye-RNA (snRNA-seq) sisebenzisa ubuchwepheshe besimanje be-10X Genomics Chromium. Le ndlela yenza banzi i-spectrum yamasampula akwazi ukuhlaziya i-transcriptome ezingeni leseli elilodwa.
Ukuhlukaniswa kwe-nuclei kufezwa nge-chip ye-10X Genomics Chromium, enohlelo lwe-microfluidics lweziteshi eziyisishiyagalombili oluwela kabili. Ngaphakathi kwalolu hlelo, ubuhlalu bejeli obuhlanganisa amabhakhodi, ama-primers, ama-enzyme, kanye ne-nucleus eyodwa kuhlanganiswe namaconsi kawoyela alingana ne-nanoliter, okwenza i-Gel Bead-in-Emulsion (GEM). Ukulandela ukwakheka kwe-GEM, i-cell lysis kanye nokukhululwa kwebhakhodi kwenzeka ngaphakathi kwe-GEM ngayinye. Kamuva, ama-molecule e-mRNA abhalwa ngokuhlehlayo abe ama-cDNA, afaka amabhakhodi angu-10X kanye Nezihlonzi Zemolekyuli Ehlukile (UMIs). Lawa ma-cDNA abe esengaphansi kokwakhiwa komtapo wolwazi wokulandelana okujwayelekile, okusiza ukuhlola okuqinile nokuphelele kwamaphrofayili wokuvezwa kofuzo ezingeni leseli elilodwa.
Inkundla: 10× Genomics Chromium kanye ne-Illumina NovaSeq Platform
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I-10x Genomics Visium Spatial Transcriptome
I-Spatial transcriptomics ubuchwepheshe obusezingeni eliphezulu obuvumela abacwaningi ukuthi baphenye amaphethini wokusho izakhi zofuzo ngaphakathi kwezicubu ngenkathi belondoloza umongo wabo wendawo. Inkundla eyodwa enamandla kulesi sizinda yi-10x Genomics Visium ehambisana nokulandelana kwe-Illumina. Umgomo we-10X Visium ulele ku-chip ekhethekile enendawo ekhethiwe yokuthwebula lapho kubekwa khona izingxenye zezicubu. Le ndawo yokuthwebula iqukethe izindawo ezinebhakhodi, ngalinye lihambisana nendawo ehlukile ngaphakathi kwethishu. Ama-molecule e-RNA athunjiwe asuka kusicubu abe eselebula ngezihlonzi eziyingqayizivele zamangqamuzana (UMIs) phakathi nenqubo yokuhlehla yokubhala. Lezi zindawo ezinebhakhodi kanye nama-UMI anika amandla ukumepha indawo okunembayo kanye nomthamo wenkulumo yofuzo ekulungisweni kweseli elilodwa. Inhlanganisela yamasampula anebhakhodi yendawo kanye nama-UMI kuqinisekisa ukunemba nokucaciswa kwedatha ekhiqizwayo. Ngokusebenzisa lobu buchwepheshe be-Spatial Transcriptomics, abacwaningi bangathola ukuqonda okujulile kwenhlangano yendawo yamangqamuzana kanye nokusebenzisana okuyinkimbinkimbi kwamangqamuzana okwenzeka ngaphakathi kwezicubu, okuhlinzeka ngemininingwane ebalulekile ezindleleni eziyisisekelo zezinqubo zebhayoloji emikhakheni eminingi, okubandakanya i-oncology, neuroscience, biology yentuthuko, i-immunology. , kanye nezifundo ze-botanical.
I-Platform: 10X Genomics Visium kanye ne-Illumina NovaSeq
