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Uburebure bwuzuye mRNA ikurikirana-Nanopore
Urutonde rwa RNA rwabaye igikoresho ntagereranywa cyo gusesengura byimazeyo.Nta gushidikanya, gakondo gakondo-isoma ikurikiranye yageze ku iterambere ryinshi hano.Nubwo bimeze bityo ariko, akenshi ihura nimbogamizi muburebure bwuzuye bwa isoform, kubara, kubogama kwa PCR.
Urutonde rwa Nanopore rwitandukanya nizindi mbuga zikurikirana, kubera ko nucleotide isomwa neza nta synthesis ya ADN kandi ikabyara igihe kirekire kuri kilobase.Ibi biha imbaraga ibyasomwe-byambukiranya inyandiko-mvugo yuzuye hamwe no gukemura ibibazo mubushakashatsi bwa isoform.
Ihuriro：Nanopore Isezerano
Isomero:cDNA-PCR
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De novo Uburebure bwuzuye Transcriptome ikurikirana -PacBio
De novobyuzuye-byuzuye bikurikirana, bizwi kandi nkaDe novoIso-Seq ifata ibyiza bya PacBio ikurikirana muburebure bwo gusoma, ituma ikurikiranwa rya molekile ya cDNA yuzuye nta kiruhuko.Ibi birinda rwose amakosa yose yatanzwe mumyandikire yinteko kandi yubaka unigene hamwe na isoform-urwego rwo gukemura.Iyi unigene igizwe itanga amakuru akomeye nka "reference genome" kuri transcriptome-urwego.Mubyongeyeho, guhuza hamwe nibisekuru bizakurikiraho bikurikirana, iyi serivise iha imbaraga umubare wuzuye wa isoform-urwego rwimvugo.
Ihuriro ： Urutonde rwa PacBio IIIsomero: Isomero ry'inzogera ya SMRT -
Eukaryotic mRNA ikurikirana-Illumina
mRNA ikurikirana ituma umwirondoro wa mRNAs wandukuwe kuva selile mubihe byihariye.Nubuhanga bukomeye bwo kwerekana imiterere ya gene, imiterere ya gene hamwe na molekuline yuburyo bumwe na bumwe bwibinyabuzima.Kugeza ubu, urutonde rwa mRNA rwakoreshejwe cyane mubushakashatsi bwibanze, gusuzuma amavuriro, guteza imbere ibiyobyabwenge, nibindi.
Ihuriro: Illumina NovaSeq 6000
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Ibidashingiye kuri mRNA ikurikirana-Illumina
mRNA ikurikirana ikoresha uburyo bukurikiraho bwo gukurikiranya (NGS) kugirango ifate intumwa RNA (mRNA) ifata Eukaryote mugihe runaka ibikorwa bimwe na bimwe bikora.Inyandiko ndende ndende yatanzwe yiswe 'Unigene' kandi ikoreshwa nk'uruhererekane rw'ibisobanuro byakurikiyeho, ubwo ni uburyo bwiza bwo kwiga uburyo bwa molekile hamwe n'urusobe rw'ibinyabuzima bitavuzwe.
Nyuma yimyandikire yamakuru hamwe na unigene ikora
(1) Isesengura rya SNP, isesengura rya SSR, guhanura CDS n'imiterere ya gene bizashyirwa ahagaragara.
(2) Kwuzuza imvugo ya unigene muri buri sample bizakorwa.
(3) Byerekanwe muburyo butandukanye hagati yintangarugero (cyangwa amatsinda) bizavumburwa hashingiwe kumvugo imwe
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Umwanya muremure utari code-Illumina
RNAs ndende idafite code (lncRNAs) ni ubwoko bwa molekile ya RNA ifite uburebure burenga 200 nt, burangwa nubushobozi buke bwa code.LncRNA, nkumunyamuryango wingenzi muri RNAs idafite code, iboneka cyane muri nucleus na plasma.Iterambere muburyo bukurikirana ikorana buhanga hamwe na bioinformtics ituma tumenya udushya twinshi lncRNAs kandi ugahuza nibikorwa byimiterere.Ibimenyetso bifatika byerekana ko lncRNA igira uruhare runini mugutegeka epigenetike, kugenzura inyandiko-mvugo no kugenzura nyuma yo kwandukura.
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Urutonde ruto rwa RNA-Illumina
RNA ntoya isobanura icyiciro cya molekile ya RNA idakunze kuba munsi ya 200nt z'uburebure, harimo micro RNA (miRNA), kwivanga kwa RNA (siRNA), hamwe na RNA (piRNA).
MicroRNA (miRNA) nicyiciro cya endogenous RNA ntoya ifite uburebure bwa 20-24nt, ikina ibintu bitandukanye byingenzi bigenga selile.miRNA igira uruhare mubikorwa byinshi byubuzima byerekana tissue - yihariye na stade - imvugo yihariye kandi ibitswe cyane mubwoko butandukanye.
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kuzenguruka RNA ikurikirana-Illumina
Urutonde rwose rwandikirwa rwashizweho kugirango rugaragaze ubwoko bwose bwa molekile ya RNA, harimo coding (mRNA) hamwe na RNAs itabigenewe (harimo lncRNA, circRNA na miRNA) byandukurwa na selile yihariye mugihe runaka.Urutonde rwose rwandikirwa, ruzwi kandi nka "RNA ikurikirana" igamije kwerekana imiyoboro yuzuye igenga urwego rwinyandiko.Kwifashisha ikoranabuhanga rya NGS, urutonde rwibicuruzwa byose byandikirwa inyandiko ziraboneka kubisesengura rya ceRNA hamwe nisesengura rya RNA, ritanga intambwe yambere iganisha kumikorere.Kugaragaza imiyoboro igenga circRNA-miRNA-mRNA ishingiye kuri ceRNA.
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Urutonde rwose rwanditse - Illumina
Urutonde rwose rwandikirwa rwashizweho kugirango rugaragaze ubwoko bwose bwa molekile ya RNA, harimo coding (mRNA) hamwe na RNAs itabigenewe (harimo lncRNA, circRNA na miRNA) byandukurwa na selile yihariye mugihe runaka.Urutonde rwose rwandikirwa, ruzwi kandi nka "RNA ikurikirana" igamije kwerekana imiyoboro yuzuye igenga urwego rwinyandiko.Kwifashisha ikoranabuhanga rya NGS, urutonde rwibicuruzwa byose byandikirwa inyandiko ziraboneka kubisesengura rya ceRNA hamwe nisesengura rya RNA, ritanga intambwe yambere iganisha kumikorere.Kugaragaza imiyoboro igenga circRNA-miRNA-mRNA ishingiye kuri ceRNA.
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Prokaryotic RNA ikurikirana
Urutonde rwa Prokaryotic RNA rukoresha ibisekuruza bizakurikiraho (NGS) kugirango bigaragaze ko RNA ihari kandi ingano mugihe runaka, ukoresheje isesengura ryimiterere ya selile.Isosiyete yacu ya prokaryotic RNA ikurikirana, igamije cyane cyane kuri prokaryote hamwe na genome yerekana, iguha ibisobanuro byerekana inyandiko-mvugo, isesengura ryimiterere ya gene, nibindi byakoreshejwe cyane mubushakashatsi bwibanze bwa siyanse, ubushakashatsi bwibiyobyabwenge niterambere, nibindi byinshi.
Ihuriro: Illumina NovaSeq 6000
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Urutonde rwa Metatranscriptome
Itondekanya rya Metatranscriptome ryerekana imiterere ya gene ya mikorobe (eukaryote na prokaryote) mubidukikije (ni ukuvuga ubutaka, amazi, inyanja, umwanda, ninda.) By'umwihariko, iyi serivise igufasha kubona imvugo ya gene yerekana imiterere ya mikorobe igoye, isesengura rya tagisi. yubwoko, isesengura ryimikorere yimikorere ya genes zitandukanye, nibindi byinshi.
Ihuriro: Illumina NovaSeq 6000
