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Sequloana se le seng sa RNA Sequencing
Tsoelo-pele ea ho nka sele e le 'ngoe le mokhoa oa ho haha laebrari ka bomong ho kopanngoa le tatellano e phahameng e lumellang lithuto tsa liphatsa tsa lefutso motheong oa sele-by-cell.E etsa hore ho be le tlhahlobo e tebileng le e felletseng ea litsamaiso tsa lisele tse rarahaneng, moo haholo-holo e qobang ho pata liphapang tsa bona ka ho nka karolelano ea lisele tsohle.
Leha ho le joalo, lisele tse ling ha li tšoanelehe ho etsoa ho emisoa ha sele e le 'ngoe, kahoo mekhoa e meng ea ho lokisa sampole - ho ntšoa ha nucleus ho tsoa liseleng hoa hlokahala, ke hore, nucleus e ntšoa ka kotloloho ho lisele kapa lisele ebe e lokisetsoa ho emisoa ha nucleus e le 'ngoe bakeng sa motho a le mong. tatellano ea lisele.
BMK e fana ka ts'ebeletso ea tatellano ea 10× Genomics ChromiumTM e thehiloeng ho sele e le 'ngoe ea RNA.Ts'ebeletso ena e sebelisitsoe haholo lithutong tse amanang le mafu, joalo ka phapang ea lisele tsa 'mele oa ho itšireletsa mafung, tumor heterogeneity, nts'etsopele ea lisele, joalo-joalo.
Spatial transcriptome chip: 10 × Genomics
Sethala: Illumina NovaSeq Platform
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BMKMANU S1000 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 hlahisitse BMKManu S1000 Spatial Transcriptome Chip, e ithorisa katharollo e ntlafalitsoengea 5µM, ho fihla ho subcellular range, le ho nolofalletsalitlhophiso tsa tharollo ea maemo a mangata.Chip ea S1000, e nang le matheba a ka bang limilione tse 2, 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 S1000 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.Tšobotsi e ikhethang ea BMKManu S1000 chip e itšetlehile ka ho feto-fetoha ha eona, e fana ka litlhophiso tsa maemo a mangata tse ka hlophisoang hantle ho lisele tse fapaneng le maemo a lintlha.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.
Ba sebelisa chip ea BMKManu S1000 le mahlale a mang a transcriptomics a sebaka, bafuputsi ba ka fumana kutloisiso e betere ea mokhatlo oa sebaka sa lisele le litšebelisano tse rarahaneng tsa limolek'hule tse etsahalang ka har'a lisele, ho fana ka leseli la bohlokoa mabapi le lits'ebetso tsa ts'ebetso ea bioloji libakeng tse fapaneng, ho kenyeletsoa. oncology, neuroscience, tsoelopele ea baeloji, immunology le lithuto tsa botanical.
Platform: BMKManu S1000 chip le Illumina NovaSeq
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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
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Bolelele bo Feletseng ba mRNA Sequencing-Nanopore
Le ha tatellano ea mRNA e thehiloeng ho NGS e sebetsa e le sesebelisoa se feto-fetohang bakeng sa palo ea polelo ea liphatsa tsa lefutso, ho itšetleha ha eona ho lipalo tse khutšoane ho thibela katleho ea eona litlhahlobong tse rarahaneng tsa transcriptomic.Nanopore sequencing, ka lehlakoreng le leng, e sebelisa theknoloji e baloang nako e telele, e nolofalletsang tatellano ea likopi tsa bolelele bo felletseng ba mRNA.Mokhoa ona o thusa ho hlahlobisisa ka botlalo mekhoa e meng ea ho arola, ho kopanya liphatsa tsa lefutso, poly-adenylation, le palo ea li-isoform tsa mRNA.
Sequencing ea Nanopore e itšetlehile ka lipontšo tsa motlakase tsa nako ea sebele tsa nanopore.E tataisoa ke liprotheine tsa enjene, DNA e nang le likhoele tse peli e hokahana le liprotheine tsa nanopore tse kentsoeng ka har'a biofilm, e phutholoha ha e feta mocha oa nanopore tlasa phapang ea voltage.Lipontšo tse ikhethang tsa motlakase tse hlahisoang ke metheo e fapaneng holim'a DNA strand li fumanoa 'me li arotsoe ka nako ea sebele, e leng ho nolofalletsang tatellano e nepahetseng le e tsoelang pele ea nucleotide.Mokhoa ona o mocha o hlola mefokolo ea ho bala nako e khuts'oane mme o fana ka sethala se matla sa tlhahlobo e rarahaneng ea genomic, e kenyelletsang lithuto tse rarahaneng tsa transcriptomic.
Sethala: Nanopore Promethion P48
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Tatelano ya bolelele bo felletseng ba mRNA -PacBio
Le ha tatellano ea mRNA e thehiloeng ho NGS e le sesebelisoa se feto-fetohang sa ho lekanya liphatsa tsa lefutso, ho itšetleha ha eona ka lipalo tse khutšoane ho thibela ts'ebeliso ea eona litlhahlobong tse rarahaneng tsa transcriptomic.PacBio sequencing (Iso-Seq), ka lehlakoreng le leng, e sebelisa theknoloji ea nako e telele e baloang, e nolofalletsang tatellano ea lingoloa tsa bolelele bo felletseng ba mRNA.Mokhoa ona o thusa ho hlahlobisisa ka mokhoa o pharaletseng oa ho arola mefuta e meng, ho kopanya liphatsa tsa lefutso le poly-adenylation le hoja e se khetho ea mantlha bakeng sa quantification ea liphatsa tsa lefutso, ka lebaka la palo e phahameng ea boitsebiso bo hlokahalang.
Theknoloji ea tatellano ea PacBio e itšetlehile ka tatellano ea molek'hule e le 'ngoe, nako ea sebele (SMRT), e fanang ka monyetla o ikhethang oa ho hapa litlaleho tsa bolelele bo feletseng ba mRNA.Mokhoa ona o mocha o kenyelletsa tšebeliso ea li-zero-mode waveguides (ZMWs), liliba tse entsoeng ka microfabricated tse nolofalletsang ho shebella nako ea sebele ea mosebetsi oa DNA polymerase nakong ea tatellano.Ka har'a li-ZMW tsena, PacBio's DNA polymerase e kopanya DNA e tlatselletsang, 'me e hlahisa lipalo tse telele tse nkang mongolo o felletseng oa mRNA.Ts'ebetso ea PacBio ka mokhoa oa Circular Consensus sequencing (CCS) e ntlafatsa ho nepahala ka ho latellana khafetsa molek'hule e tšoanang.Libalo tsa HiFi tse hlahisitsoeng li na le ho nepahala ho ka bapisoang le NGS, tse tlatsetsang hape tlhahlobong e felletseng le e tšepahalang ea likarolo tse rarahaneng tsa transcriptomic.Sethala: PacBio Sequel II
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Eukaryotic mRNA Sequencing-Illumina
tatellano ea mRNA e matlafatsa profil e felletseng ea lingoliloeng tsohle tsa mRNA ka har'a lisele tlas'a maemo a ikhethileng.Theknoloji ena e tsoetseng pele e sebetsa e le sesebelisoa se matla, se senolang liphatlalatso tse rarahaneng tsa liphatsa tsa lefutso, libopeho tsa liphatsa tsa lefutso, le mekhoa ea limolek'hule e amanang le mekhoa e fapaneng ea tlhaho.E amohetsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, le nts'etsopele ea lithethefatsi, tatellano ea mRNA e fana ka leseli mabapi le ho rarahana ha matla a cellular le taolo ea liphatsa tsa lefutso.
Sethala: Illumina NovaSeq X
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Non-Reference based mRNA Sequencing-Illumina
tatellano ea mRNA e matlafatsa profil e felletseng ea lingoliloeng tsohle tsa mRNA ka har'a lisele tlas'a maemo a ikhethileng.Theknoloji ena e tsoetseng pele e sebetsa e le sesebelisoa se matla, se senolang liphatlalatso tse rarahaneng tsa liphatsa tsa lefutso, libopeho tsa liphatsa tsa lefutso, le mekhoa ea limolek'hule e amanang le mekhoa e fapaneng ea tlhaho.E amohetsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, le nts'etsopele ea lithethefatsi, tatellano ea mRNA e fana ka leseli mabapi le ho rarahana ha matla a cellular le taolo ea liphatsa tsa lefutso.
Sethala: Illumina NovaSeq X
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Tatelano ya nako e telele e sa khoutong-Illumina
Li-RNA tse telele tse sa khouteng (lncRNAs) ke li-RNA tse telele ho feta li-nucleotide tse 200 tse nang le bokhoni bo fokolang ba ho khouta ebile e le likarolo tsa bohlokoa ka har'a RNA e sa khouteng.E fumaneha nucleus le cytoplasm, li-RNA tsena li bapala karolo ea bohlokoa taolong ea epigenetic, transcriptal, le post-transcriptal, e totobatsang bohlokoa ba tsona ho bopeng lits'ebetso tsa cellular le limolek'hule.LncRNA sequencing ke sesebelisoa se matla ho arola lisele, Ontogenesis, le mafu a Batho.
Sethala: Illumina NovaSeq
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Nyenyane RNA sequencing-Illumina
Limolek'hule tse nyane tsa RNA (sRNA), hangata tse ka tlase ho li-nucleotide tse 200 ka bolelele, li kenyelletsa li-microRNAs (miRNAs), li-RNA tse kenang tse nyane (siRNAs), le li-RNA tse sebetsanang le piwi (piRNAs).Har'a tsena, li-miRNA, tse ka bang bolelele ba 20-24 nucleotide, li bohlokoa haholo bakeng sa likarolo tsa tsona tsa bohlokoa tsa taolo lits'ebetsong tse fapaneng tsa cellular.Ka mekhoa e ikhethileng ea lisele le sethala, miRNA e bonts'a paballo e phahameng ho pholletsa le mefuta e fapaneng.
Sethala: Illumina NovaSeq
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circRNA sequencing-Illumina
Circry RNA sequencing (circRNA-seq) ke ho hlahisa le ho hlahloba li-RNA tse chitja, sehlopha sa limolek'hule tsa RNA tse etsang li-loops tse koetsoeng ka lebaka la liketsahalo tse sa lumellaneng le li-canonical, ho fana ka RNA tsena ka botsitso bo eketsehileng.Le ha li-circRNA tse ling li bonts'itsoe hore li sebetsa joalo ka liponche tse nyane tsa microRNA, li ts'oara li-microRNA le ho li thibela ho laola li-mRNA tsa tsona, li-circRNA tse ling li ka sebelisana le liprotheine, tsa fetola liphatsa tsa lefutso, kapa tsa ba le likarolo lits'ebetsong tsa cellular.Tlhahlobo ea polelo ea circRNA e fana ka leseli mabapi le likarolo tsa taolo ea limolek'hule tsena le bohlokoa ba tsona mekhoeng e fapaneng ea lisele, methati ea nts'etsopele, le maemo a mafu, e tlatsetsang kutloisisong e tebileng ea ho rarahana ha taolo ea RNA maemong a polelo ea lefutso.
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Tatelano e felletseng ea transcriptome - Illumina
Tatelano e felletseng ea li-transcriptome e fana ka mokhoa o felletseng oa ho hlahisa limolek'hule tse fapaneng tsa RNA, tse kenyelletsang li-coding (mRNA) le li-RNA tse sa khoutang (lncRNA, circRNA, le miRNA).Mokhoa ona o nka transcriptome e felletseng ea lisele tse itseng ka nako e itseng, e lumellang kutloisiso e felletseng ea lits'ebetso tsa cellular.E boetse e tsejoa e le "total RNA sequencing," e ikemiselitse ho senola marang-rang a rarahaneng a taolo boemong ba transcriptome, ho nolofalletsa tlhahlobo e tebileng joalo ka tlholisano ea endogenous RNA (ceRNA) le tlhahlobo e kopaneng ea RNA.Sena se tšoaea mohato oa pele o lebisang ho sebopeho se sebetsang, haholo-holo ho manolleng marang-rang a taolo a kenyelletsang litšebelisano tsa ceRNA tse thehiloeng ho circRNA-miRNA-mRNA.
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Prokaryotic mRNA tatelano
tatellano ea mRNA e matlafatsa profil e felletseng ea lingoliloeng tsohle tsa mRNA ka har'a lisele tlas'a maemo a ikhethileng.Theknoloji ena e tsoetseng pele e sebetsa e le sesebelisoa se matla, se senolang liphatlalatso tse rarahaneng tsa liphatsa tsa lefutso, libopeho tsa liphatsa tsa lefutso, le mekhoa ea limolek'hule e amanang le mekhoa e fapaneng ea tlhaho.E amohetsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, le nts'etsopele ea lithethefatsi, tatellano ea mRNA e fana ka leseli mabapi le ho rarahana ha matla a cellular le taolo ea liphatsa tsa lefutso.Ts'ebetso ea rona ea sampole ea prokaryotic mRNA e etselitsoe li-transcriptomes tsa prokaryotic, tse kenyelletsang ho fokotseha ha rRNA le ho hlophisoa ha laeborari ea tataiso.
Sethala: Illumina NovaSeq X
