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BMKMANU S3000_Spatial Transcriptome
Spatial transcriptomics ndi njira yomwe imatilola kujambula ndikuwona mawonekedwe a jini mkati mwa minofu. Izi zitha kukhala zofunikira kuti timvetsetse momwe ma cell amalumikizirana.
Pali nsanja zosiyanasiyana za njira iyi. Pankhaniyi, BMKGene yapanga BMKManu 3000 Spatial transcriptome Chip, nsanja yomwe imakulitsa luso laukadaulo, kufikira kusamvana kwa ma cell ndikuthandizira kukhazikika kwamagawo angapo.
Chip ichi chimatsekera mawanga 4.2 miliyoni pogwiritsa ntchito ukadaulo wovomerezeka wa tinthu tating'onoting'ono tomwe timakhala ndi mikanda yodzaza ndi ma probe okhala ndi barcode. Ndi njira iyi, pambuyo kulanda ndi kukulitsa, timapeza laibulale ya cDNA yolemeretsedwa ndi zitsanzo za barcode zomwe ndi Illumina yogwirizana.
Pazidziwitso, kuphatikiza kwa barcode ya malo ndi UMIs kumatsimikizira kulondola ndi tsatanetsatane wa deta yopangidwa. Kuphatikiza zonse zomwe tafotokozazi, BMKManu imapereka mawonekedwe osunthika kwambiri.
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Single-nuclei RNA Sequencing
Kupanga makina ojambulira selo limodzi ndi njira zomangira laibulale yanthawi zonse, kuphatikizidwa ndi kutsatizana kopitilira muyeso, kwasintha maphunziro a jini pamaselo. Kupambana kumeneku kumapangitsa kusanthula mozama komanso mozama za kuchuluka kwa maselo ovuta, kuthana ndi malire omwe amakhudzana ndi kuchuluka kwa ma jini pama cell onse ndikusunga kusiyanasiyana kowona m'maguluwa. Ngakhale kutsatizana kwa selo imodzi ya RNA (scRNA-seq) kuli ndi ubwino wosatsutsika, kumakumana ndi zovuta m'magulu ena kumene kupanga kuyimitsidwa kwa selo limodzi kumakhala kovuta ndipo kumafuna zitsanzo zatsopano. Ku BMKGene, timathana ndi vutoli popereka nyukiliya imodzi ya RNA sequencing (snRNA-seq) pogwiritsa ntchito ukadaulo wapamwamba kwambiri wa 10X Genomics Chromium. Njirayi imakulitsa kuchuluka kwa zitsanzo zomwe zimatha kusanthula transcriptome pamlingo wa selo limodzi.
Kudzipatula kwa ma nuclei kumachitika kudzera mu chipangizo chamakono cha 10X Genomics Chromium, chokhala ndi makina asanu ndi atatu a microfluidics okhala ndi kuwoloka kawiri. Mkati mwa dongosololi, mikanda ya gel yomwe imaphatikizapo ma barcode, zoyambira, ma enzyme, ndi phata limodzi zimayikidwa mu madontho amafuta a nanoliter, kupanga Gel Bead-in-Emulsion (GEM). Kutsatira kupangidwa kwa GEM, cell lysis ndi barcode kumasulidwa kumachitika mkati mwa GEM iliyonse. Pambuyo pake, mamolekyu a mRNA amalembedwa m'ma cDNAs, kuphatikiza ma barcode 10X ndi Unique Molecular Identifiers (UMIs). Ma cDNA awa amasinthidwa kukhala laibulale yotsatizana, kuthandizira kuwunika kozama komanso kozama kwa mbiri ya jini pamlingo wa selo imodzi.
Pulatifomu: 10 × Genomics Chromium ndi Illumina NovaSeq Platform
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10x Genomics Visium Spatial Transcriptome
Spatial transcriptomics ndiukadaulo wotsogola womwe umalola ofufuza kuti afufuze momwe ma jini amafotokozera mkati mwa minofu ndikusunga malo awo. Pulatifomu imodzi yamphamvu mderali ndi 10x Genomics Visium yophatikizidwa ndi kutsatizana kwa Illumina. Mfundo ya 10X Visium yagona pa chip chapadera chokhala ndi malo osankhidwa omwe magawo a minofu amayikidwa. Malo ogwidwawa ali ndi mabala a barcode, omwe amafanana ndi malo apadera mkati mwa minofu. Mamolekyu a RNA omwe adagwidwa kuchokera ku minofuyo amalembedwa ndi zozindikiritsa za mamolekyulu (UMIs) panthawi yolemba. Malo okhala ndi barcode ndi ma UMI amathandizira kupanga mapu olondola a malo komanso kuchuluka kwa mafotokozedwe a jini pakusintha kwa selo limodzi. Kuphatikiza kwa zitsanzo zokhala ndi barcode ndi UMIs zimatsimikizira kulondola ndi kutsimikizika kwa deta yopangidwa. Pogwiritsa ntchito ukadaulo wa Spatial Transcriptomics, ofufuza atha kumvetsetsa mozama za momwe ma cell amapangidwira komanso momwe ma cell amagwirira ntchito mkati mwa minyewa, ndikupereka chidziwitso chamtengo wapatali pamakina omwe amachitika m'magawo angapo, kuphatikiza oncology, neuroscience, biology yachitukuko, immunology, ndi maphunziro a botanical.
Platform: 10X Genomics Visium ndi Illumina NovaSeq
