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BMKMANU S3000_Spatial Transcriptome
Li-Spatial transcriptomics li ka pele ho boqapi ba mahlale, li matlafatsa bafuputsi ho batlisisa mekhoa e rarahaneng ea liphatsa tsa lefutso ka har'a lisele ha ba ntse ba boloka maemo a bona a sebaka. Har'a li-platform tse fapaneng, BMKGene e ntlafalitse BMKManu S3000 Spatial Transcriptome Chip, e ithorisa ka qeto e ntlafalitsoeng ea 3.5µm, e fihlang ho subcellular range, le ho nolofalletsa litlhophiso tsa tharollo ea maemo a mangata. Chip ea S3000, e nang le matheba a ka bang limilione tse 4, e sebelisa li-microwell tse koahetsoeng ka lifaha tse nang le lisebelisoa tsa ho hapa tse nang le barcode. Laeborari ea cDNA, e ntlafalitsoeng ka li-barcode tsa sebaka, e lokisoa ho tsoa ho chip ea S3000 mme ka mor'a moo e hlophisoa sethaleng sa Illumina NovaSeq. Motsoako oa disampole tse nang le barcoded tsa sebaka le li-UMI li netefatsa ho nepahala le ho nepahala ha data e hlahisitsoeng. The BMKManu S3000 chip's e sebetsa ka mokhoa o fapaneng haholo, e fana ka litlhophiso tsa maemo a mangata tse ka hlophisoang hantle le lisele tse fapaneng le lintlha tse batloang. Ho ikamahanya le maemo hona ho beha chip e le khetho e ikhethang bakeng sa lithuto tse fapaneng tsa transcriptomics tsa sebaka, e netefatsang kopano e nepahetseng ea sebaka ka lerata le fokolang. Ts'ebeliso ea mahlale a ho arola lisele ka BMKManu S3000 e thusa ho aroloa ha data ea mongolo ho meeli ea lisele, e leng se hlahisang tlhahlobo e nang le moelelo o tobileng oa tlhaho. Ho feta moo, qeto e ntlafetseng ea S3000 e fella ka palo e phahameng ea liphatsa tsa lefutso le li-UMI tse fumanoang ka sele, e leng ho nolofalletsang tlhahlobo e nepahetseng haholoanyane ea mekhoa ea ho ngola sebaka le ho bokellana ha lisele.
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Single-nuclei RNA Sequencing
Nts'etsopele ea ho ts'oaroa ha sele e le 'ngoe le mekhoa ea kaho ea laeborari e tloahelehileng, hammoho le tatellano ea maemo a holimo, e fetotse lithuto tsa liphatsa tsa lefutso maemong a lisele. Katleho ena e lumella tlhahlobo e tebileng le e felletseng ea lisele tse rarahaneng tsa lisele, ho hlola mefokolo e amanang le karolelano ea liphatsa tsa lefutso holim'a lisele tsohle le ho boloka ho fapana ha 'nete ka har'a batho bana. Le ha tatellano ea sele e le 'ngoe ea RNA (scRNA-seq) e na le melemo e ke keng ea latoloa, e kopana le liqholotso meleng e itseng moo ho theoang ho emisoa ha sele e le 'ngoe ho ipakang ho le thata mme ho hloka lisampole tse ncha. Ho BMKGene, re rarolla bothata bona ka ho fana ka tatellano ea "nucleus RNA" e le 'ngoe (snRNA-seq) re sebelisa theknoloji ea morao-rao ea 10X Genomics Chromium. Mokhoa ona o pharalatsa mefuta e mengata ea lisampole tse khonang ho hlahloba transcriptome boemong ba sele e le 'ngoe.
Ho aroloa ha li-nuclei ho finyelloa ka chip ea 10X Genomics Chromium e ncha, e nang le tsamaiso ea microfluidics e nang le mechine e robeli e nang le litsela tse peli. Ka har'a tsamaiso ena, lifaha tsa li-gel tse kenyelletsang li-barcode, li-primers, li-enzyme, le nucleus e le 'ngoe li kenngoa ka marotholi a oli a boholo ba nanoliter, ho etsa Gel Bead-in-Emulsion (GEM). Ka mor'a ho thehoa ha GEM, lisele tsa lysis le ho lokolloa ha barcode ho etsahala ka har'a GEM ka 'ngoe. Ka mor'a moo, limolek'hule tsa mRNA li fetoleloa ka morao ho li-cDNA, tse kenyelletsang li-barcode tsa 10X le Unique Molecular Identifiers (UMIs). Li-cDNA tsena li beoa tlas'a kaho e tloaelehileng ea tatellano ea laeborari, ho thusa tlhahlobo e matla le e felletseng ea liphatlalatso tsa liphatsa tsa lefutso boemong ba sele e le 'ngoe.
Sethala: 10× Genomics Chromium le Illumina NovaSeq Platform
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10x Genomics Visium Spatial Transcriptome
Spatial transcriptomics ke theknoloji e tsoetseng pele e lumellang bafuputsi ho etsa lipatlisiso tsa mefuta ea liphatsa tsa lefutso ka har'a lisele ha ba ntse ba boloka maemo a bona a sebaka. Sethala se le seng se matla sebakeng sena ke 10x Genomics Visium e kopantsoeng le tatellano ea Illumina. Molao-motheo oa 10X Visium o holim'a chip e khethehileng e nang le sebaka se khethiloeng sa ho hapa moo likarolo tsa lisele li behoang teng. Sebaka sena sa ho ts'oaroa se na le libaka tse nang le barcode, e 'ngoe le e' ngoe e tsamaellana le sebaka se ikhethileng ka har'a thishu. Limolek'hule tsa RNA tse hapuoeng ho tsoa liseleng ebe li ngotsoe ka li-identifiers tse ikhethang tsa limolek'hule (UMIs) nakong ea ts'ebetso ea ho fetolela ka morao. Libaka tsena tse nang le li-barcode le li-UMI li thusa ho etsa 'mapa o nepahetseng oa sebaka le ho lekanya liphatsa tsa lefutso ka qeto ea sele e le 'ngoe. Motsoako oa disampole tse nang le barcoded tsa sebaka le li-UMI li netefatsa ho nepahala le ho nepahala ha data e hlahisitsoeng. Ka ho sebelisa theknoloji ena ea Spatial Transcriptomics, bafuputsi ba ka fumana kutloisiso e tebileng ea mokhatlo oa sebaka sa lisele le litšebelisano tse rarahaneng tsa limolek'hule tse hlahang ka har'a lisele, tse fanang ka kutloisiso ea bohlokoa ka mekhoa ea mekhoa ea likokoana-hloko likarolong tse ngata, ho akarelletsa le oncology, neuroscience, tsoelo-pele ea biology, immunology. , le lithuto tsa botanical.
Sethala: 10X Genomics Visium le Illumina NovaSeq
