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Tatelano ya bolelele bo felletseng ba mRNA-Nanopore
Tatelano ea RNA e bile sesebelisoa sa bohlokoahali bakeng sa tlhahlobo e felletseng ea transcriptome.Ntle le pelaelo, tatelano ea khale ea ho bala nako e khuts'oane e fihletse tsoelo-pele e mengata ea bohlokoa mona.Leha ho le joalo, hangata e kopana le mefokolo ho li-isoform tsa bolelele bo felletseng, quantification, leeme la PCR.
Nanopore sequencing e ikhetholla ho li-platform tse ling tse latellanang, ka hore li-nucleotide li baloa ka ho toba ntle le motsoako oa DNA 'me li hlahisa nako e telele ho bala li-kilobase tse mashome.Sena se fana ka matla a ho bala lingoliloeng tsa bolelele bo felletseng le ho sebetsana le liphephetso lithutong tsa boemo ba isoform.
Sethala:Nanopore PromethION
Laeborari:cDNA-PCR
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De novo Full-length Transcriptome sequencing -PacBio
Ho fihlela joaletatellano ea bolelele bo felletseng ba transcriptome, e tsejoang hape e leHo fihlela joaleIso-Seq e nka melemo ea sequencer ea PacBio ka bolelele ba ho bala, e nolofalletsang tatellano ea limolek'hule tsa cDNA tsa bolelele bo felletseng ntle le khefu.Sena se qoba ka ho feletseng liphoso leha e le life tse hlahisoang mehatong ea kopano e ngotsoeng 'me se haha sete sa unigene ka qeto ea boemo ba isoform.Sehlopha sena sa unigene se fana ka tlhahisoleseding e matla ea liphatsa tsa lefutso e le "reference genome" ho transcriptome-level.Ntle le moo, ho kopantsoe le data ea tatellano ea moloko o latelang, ts'ebeletso ena e matlafatsa quantification e nepahetseng ea polelo ea boemo ba isoform.
Sethala: PacBio Sequel IILaebrari: Laeborari ea tšepe ea SMRT -
Eukaryotic mRNA sequencing-Illumina
tatellano ea mRNA e thusa ho fana ka profil ea li-mRNA tsohle tse ngotsoeng ho tsoa liseleng tlas'a maemo a itseng.Ke thekenoloji e matla ea ho senola boemo ba liphatsa tsa lefutso, mekhoa ea liphatsa tsa lefutso le mekhoa ea limolek'hule ea mekhoa e itseng ea likokoana-hloko.Ho fihla hajoale, tatellano ea mRNA e sebelisitsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, nts'etsopele ea lithethefatsi, jj.
Sethala: Illumina NovaSeq 6000
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Non-Reference based mRNA sequencing-Illumina
tatellano ea mRNA e sebelisa mokhoa oa tatellano oa moloko o latelang (NGS) ho hapa lenģosa RNA(mRNA) foromo ea Eukaryote ka nako e itseng eo mesebetsi e meng e khethehileng e ntseng e sebetsa.Sengoliloeng se selelele ka ho fetesisa se ne se bitsoa 'Unigene' 'me se sebelisitsoe e le tatelano ea litšupiso bakeng sa tlhahlobo e latelang, e leng mokhoa o sebetsang oa ho ithuta mokhoa oa limolek'hule le marang-rang a taolo ea mefuta ntle le ho buuoa.
Ka mor'a kopano ea data ea transcriptome le tlhaloso e sebetsang ea unigene
(1) Tlhahlobo ea SNP, tlhahlobo ea SSR, ho bolela esale pele ka CDS le sebopeho sa liphatsa tsa lefutso li tla etsoa esale pele.
(2) Quantification ea unigene expression ho sampole ka 'ngoe e tla etsoa.
(3) Li-unigene tse hlahisitsoeng ka mokhoa o fapaneng lipakeng tsa lisampole (kapa lihlopha) li tla fumanoa ho ipapisitsoe le polelo e fapaneng.
(4) Ho tla etsoa tlhahlobo ea lihlopha, tlhaloso e sebetsang le tlhahlobo ea ntlafatso ea li-unigenes tse hlalositsoeng ka tsela e fapaneng.
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Nako e telele e sa kh'outu ea tatelano-Illumina
Li-RNA tse telele tse sa khouteng (lncRNAs) ke mofuta oa limolek'hule tsa RNA tse bolelele bo fetang 200 nt, tse nang le bokhoni bo tlase haholo ba ho khouta.LncRNA, joalo ka setho se ka sehloohong ho li-RNA tse sa ngoliseng likhoutu, e fumaneha haholo ka har'a nucleus le plasma.Nts'etsopele ea tatellano ea theknoloji le bioinformatics e thusa ho tsebahatsa li-lncRNA tse ngata le ho amahanya tse nang le mesebetsi ea tlhaho.Bopaki bo bokelletsoeng bo fana ka maikutlo a hore lncRNA e ameha haholo ho epigenetic regulation, transcription regulation le post-transcription regulation.
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Nyenyane RNA sequencing-Illumina
RNA e nyane e bua ka sehlopha sa limolek'hule tsa RNA tse sa khoute tseo hangata li leng ka tlase ho 200nt ka bolelele, ho kenyeletsoa RNA e nyane (miRNA), tšitiso e nyane ea RNA (siRNA), le RNA e sebetsanang le piwi (piRNA).
MicroRNA (miRNA) ke sehlopha sa RNA e nyane ea endogenous e bolelele ba hoo e ka bang 20-24nt, e bapalang likarolo tse fapaneng tsa taolo ea lisele.miRNA e kenya letsoho lits'ebetsong tse ngata tsa bophelo tse senolang lisele - tse ikhethang le tsa sethala - polelo e ikhethileng le e bolokiloeng haholo mefuteng e fapaneng.
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circRNA sequencing-Illumina
Whole transcriptome sequencing e etselitsoe ho profil mefuta eohle ea limolek'hule tsa RNA, ho kenyeletsoa coding (mRNA) le li-RNA tse sa khoute (ho kenyeletsoa lncRNA, circRNA le miRNA) tse ngotsoeng ke lisele tse itseng ka nako e itseng.Whole transcriptome sequencing, e tsejoang hape e le "total RNA sequencing" e ikemiselitse ho senola marang-rang a felletseng a taolo maemong a transcriptome.Ho nka monyetla ka theknoloji ea NGS, tatellano ea lihlahisoa tsohle tsa transcriptome lia fumaneha bakeng sa tlhahlobo ea ceRNA le tlhahlobo e kopaneng ea RNA, e fanang ka mohato oa pele oa ho hlahisa sebopeho se sebetsang.Ho senola marang-rang a taolo ea ceRNA e thehiloeng ho circRNA-miRNA-mRNA.
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Tatelano e felletseng ea transcriptome - Illumina
Whole transcriptome sequencing e etselitsoe ho profil mefuta eohle ea limolek'hule tsa RNA, ho kenyeletsoa coding (mRNA) le li-RNA tse sa khoute (ho kenyeletsoa lncRNA, circRNA le miRNA) tse ngotsoeng ke lisele tse itseng ka nako e itseng.Whole transcriptome sequencing, e tsejoang hape e le "total RNA sequencing" e ikemiselitse ho senola marang-rang a felletseng a taolo maemong a transcriptome.Ho nka monyetla ka theknoloji ea NGS, tatellano ea lihlahisoa tsohle tsa transcriptome lia fumaneha bakeng sa tlhahlobo ea ceRNA le tlhahlobo e kopaneng ea RNA, e fanang ka mohato oa pele oa ho hlahisa sebopeho se sebetsang.Ho senola marang-rang a taolo ea ceRNA e thehiloeng ho circRNA-miRNA-mRNA.
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Prokaryotic RNA tatelano
Prokaryotic RNA sequencing e sebelisa tatellano ea moloko o latelang (NGS) ho senola boteng le bongata ba RNA ka nako e itseng, ka ho hlahloba phetoho ea cellular transcriptome.K'hamphani ea rona ea prokaryotic RNA sequencing, e ikemiselitse haholo ho li-prokaryotes tse nang le liphatsa tsa lefutso, ho u fa profiling ea transcriptome, tlhahlobo ea sebopeho sa liphatsa tsa lefutso, joalo-joalo. E sebelisitsoe haholo lipatlisisong tsa mahlale a mantlha, lipatlisiso tsa lithethefatsi le nts'etsopele, le tse ling.
Sethala: Illumina NovaSeq 6000
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Metatranscriptome Sequencing
Metatranscriptome sequencing e supa liphatsa tsa lefutso tsa likokoana-hloko (ka bobeli eukaryotes le prokaryotes) ka har'a tikoloho ea tlhaho (e leng mobu, metsi, leoatle, mantle le mala.) ea mefuta, tlhahlobo ea ntlafatso ea ts'ebetso ea liphatsa tsa lefutso tse hlahisitsoeng ka mokhoa o fapaneng, le tse ling.
Sethala: Illumina NovaSeq 6000
