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ʻO ke kaʻina mRNA holoʻokoʻa-Nanopore
He mea waiwai nui ka RNA sequencing no ka loiloi transcriptome piha.ʻO ka mea kānalua, ua hoʻokō ʻia nā hoʻomohala koʻikoʻi o ka heluhelu pōkole kuʻuna ma ʻaneʻi.Eia nō naʻe, ʻike pinepine ia i nā palena i nā ʻike isoform holoʻokoʻa, quantification, PCR bias.
Hoʻokaʻawale ʻo Nanopore sequencing iā ia iho mai nā paepae hoʻonohonoho ʻē aʻe, no ka mea, heluhelu pololei ʻia nā nucleotides me ka ʻole o ka DNA synthesis a hoʻopuka i ka heluhelu lōʻihi ma nā ʻumi kilobases.Hoʻoikaika kēia i ka heluhelu ʻana i nā transcript lōʻihi piha a me ka hoʻoponopono ʻana i nā pilikia i nā haʻawina isoform-level.
Papahana：Nanopore PromethION
Hale Waihona Puke:cDNA-PCR
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ʻO De novo Ke kaʻina hana Transcriptome holoʻokoʻa -PacBio
De novoka lōʻihi transcriptome sequencing, i ʻike ʻia ʻoDe novoLawe ʻo Iso-Seq i nā pono o ka Sequencer PacBio i ka lōʻihi heluhelu, e hiki ai i ka hoʻonohonoho ʻana i nā molekala cDNA piha me ka ʻole o ka haki.Hōʻalo loa kēia i nā hewa i hana ʻia ma nā ʻanuʻu hui transcript a kūkulu i nā pūʻulu unigene me ka hoʻonā isoform-level.Hāʻawi kēia unigene set i ka ʻike genetic ikaika e like me "genome reference" ma ka pae transcriptome.Eia kekahi, hui pū me ka ʻikepili sequencing o ka hanauna e hiki mai ana, hāʻawi kēia lawelawe i ka helu kūpono o ka ʻōlelo isoform-level.
Papahana: PacBio Sequel IIHale Waihona Puke: SMRT bele waihona -
Eukaryotic mRNA sequencing-Illumina
Hiki i ka mRNA sequencing ke hoʻopili i nā mRNA āpau i unuhi ʻia mai nā cell ma lalo o nā kūlana kikoʻī.He ʻenehana ikaika ia no ka hōʻike ʻana i ka moʻolelo hōʻike gene, nā ʻano hana a me nā hana molekala o kekahi mau kaʻina olaola.I kēia lā, ua hoʻohana nui ʻia ka mRNA sequencing i ka noiʻi kumu, diagnostics clinical, development drug, etc.
Papahana: Illumina NovaSeq 6000
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Non-Reference based mRNA sequencing-Illumina
Hoʻohana ʻo mRNA sequencing i ka ʻenehana sequencing hanauna hou (NGS) e hopu i ka messenger RNA(mRNA) form Eukaryote i ka manawa kikoʻī e hoʻāla ana kekahi mau hana kūikawā.ʻO ka transcript lōʻihi loa i hoʻopili ʻia ua kapa ʻia ʻo 'Unigene' a hoʻohana ʻia ma ke ʻano he kaʻina kuhikuhi no ka nānā ʻana ma hope, ʻo ia ke ala kūpono e aʻo ai i ka mīkini molekala a me ka ʻoihana hoʻoponopono o nā ʻano me ka ʻole o ka kuhikuhi.
Ma hope o ka hōʻuluʻulu ʻana o ka ʻikepili transcriptome a me ka annotation hana unigene
(1) E hoʻonohonoho mua ʻia ka ʻike SNP, ka nānā ʻana SSR, ka wānana CDS a me ka hoʻolālā gene.
(2) E hana ʻia ka helu o ka ʻōlelo unigene i kēlā me kēia laʻana.
(3) E ʻike ʻia nā unigenes i hōʻike ʻia ma waena o nā laʻana (a i ʻole nā hui) ma muli o ka ʻōlelo unigene.
(4) E hoʻokō ʻia ka Clustering, ka wehewehe hana a me ka hoʻonui ʻana i nā unigenes i hōʻike ʻia.
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Ka lōʻihi non-coding sequencing-Illumina
ʻO nā RNA non-coding lōʻihi (lncRNAs) he ʻano molekole RNA me ka lōʻihi ma mua o 200 nt, kahi i hōʻike ʻia e ka haʻahaʻa haʻahaʻa loa.ʻO LncRNA, ma ke ʻano he lālā koʻikoʻi i nā RNA non-coding, ʻike nui ʻia i ka nucleus a me ka plasma.ʻO ka hoʻomohala ʻana i ka ʻenehana sequencing a me bioinformtics hiki ke ʻike i nā lncRNA hou a hoʻopili i nā mea me nā hana biological.Hōʻike nā hōʻike hōʻuluʻulu e pili nui ana ka lncRNA i ka hoʻoponopono epigenetic, ka hoʻoponopono transcription a me ka hoʻoponopono post-transcription.
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Liʻiliʻi RNA sequencing-Illumina
ʻO ka RNA liʻiliʻi e pili ana i kahi papa o nā molekole RNA non-coding i emi iho ma mua o 200nt ka lōʻihi, me ka micro RNA (miRNA), RNA interference liʻiliʻi (siRNA), a me ka piwi-interacting RNA (piRNA).
ʻO MicroRNA (miRNA) kahi papa o RNA liʻiliʻi endogenous me ka lōʻihi o 20-24nt, e pāʻani ana i nā ʻano koʻikoʻi koʻikoʻi koʻikoʻi i loko o nā cell.Hoʻokomo ʻia ka miRNA i nā kaʻina ola he nui e hōʻike ana i nā kiko - kikoʻī a me ka pae - ʻōlelo kikoʻī a mālama nui ʻia i nā ʻano ʻano like ʻole.
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circRNA sequencing-Illumina
Hoʻolālā ʻia ke kaʻina transcriptome holoʻokoʻa e hōʻike i nā ʻano molekole RNA āpau, me ka coding (mRNA) a me nā RNA non-coding (me lncRNA, circRNA a me miRNA) i kākau ʻia e nā cell kikoʻī i kekahi manawa.ʻO ke kaʻina transcriptome holoʻokoʻa, i ʻike ʻia ʻo "total RNA sequencing" e manaʻo e hōʻike i nā ʻoihana hoʻoponopono piha i ka pae transcriptome.ʻO ka hoʻohana ʻana i ka ʻenehana NGS, loaʻa nā kaʻina o nā huahana transcriptome holoʻokoʻa no ka nānā ʻana ceRNA a me ka loiloi RNA hui, e hāʻawi ana i ka hana mua i ka ʻano hana.Hōʻike i ka pūnaewele hoʻoponopono o circRNA-miRNA-mRNA e pili ana i ka ceRNA.
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ʻO ke kaʻina transcriptome holoʻokoʻa - Illumina
Hoʻolālā ʻia ke kaʻina transcriptome holoʻokoʻa e hōʻike i nā ʻano molekole RNA āpau, me ka coding (mRNA) a me nā RNA non-coding (me lncRNA, circRNA a me miRNA) i kākau ʻia e nā cell kikoʻī i kekahi manawa.ʻO ke kaʻina transcriptome holoʻokoʻa, i ʻike ʻia ʻo "total RNA sequencing" e manaʻo e hōʻike i nā ʻoihana hoʻoponopono piha i ka pae transcriptome.ʻO ka hoʻohana ʻana i ka ʻenehana NGS, loaʻa nā kaʻina o nā huahana transcriptome holoʻokoʻa no ka nānā ʻana ceRNA a me ka loiloi RNA hui, e hāʻawi ana i ka hana mua i ka ʻano hana.Hōʻike i ka pūnaewele hoʻoponopono o circRNA-miRNA-mRNA e pili ana i ka ceRNA.
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Ka hoʻokaʻina RNA Prokaryotic
Hoʻohana ka prokaryotic RNA sequencing i ka hanauna e hiki mai ana (NGS) e hōʻike i ka hele ʻana a me ka nui o RNA i ka manawa i hāʻawi ʻia, ma ke kāʻili ʻana i ka transcriptome cellular loli.ʻO ka prokaryotic RNA sequencing o kā mākou hui, pili pono i nā prokaryotes me nā genomes reference, hāʻawi iā ʻoe me ka transcriptome profiling, gene structure analysis, etc.
Papahana: Illumina NovaSeq 6000
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Hoʻokaʻina Metatranscriptome
ʻO ka metatranscriptome sequencing e hōʻike ana i ka hōʻike gene o nā microbes (ʻo nā eukaryotes a me nā prokaryotes) i loko o nā wahi kūlohelohe (ʻo ia hoʻi ka lepo, ka wai, ke kai, nā feces, a me ka ʻōpū.). o nā ʻano, ka loiloi hoʻonui hana o nā ʻano genes i hōʻike ʻia, a me nā mea hou aku.
Papahana: Illumina NovaSeq 6000
