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Tag nrho-ntev mRNA sequencing-Nanopore
RNA sequencing yog ib qho cuab yeej tseem ceeb rau kev txheeb xyuas cov ntaub ntawv sau tseg.Doubtlessly, ib txwm nyeem luv luv sequencing ua tiav ntau txoj kev loj hlob tseem ceeb hauv no.Txawm li cas los xij, nws feem ntau ntsib cov kev txwv nyob rau hauv tag nrho-ntev isoform identifications, quantification, PCR bias.
Nanopore sequencing distinguishes nws tus kheej los ntawm lwm cov sequencing platforms, nyob rau hauv hais tias cov nucleotides yog nyeem ncaj qha tsis muaj DNA synthesis thiab generates ntev nyeem ntawm kaum ntawm kilobases.Qhov no txhawb nqa kev nyeem ntawv ncaj qha hla cov ntawv teev lus ntev ntev thiab daws cov teeb meem hauv isoform-theem kev tshawb fawb.
Platform:Nanopor PromethION
Lub tsev qiv ntawv:cDNA-PCR
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De novo Full-length Transscriptome sequencing -PacBio
De novfull-length transcriptome sequencing, tseem hu uaDe novIso-Seq siv qhov zoo ntawm PacBio sequencer hauv kev nyeem ntev, uas ua rau kev sib txuas ntawm tag nrho-ntev cDNA molecules yam tsis muaj kev so.Qhov no ua kom tsis txhob muaj qhov yuam kev tsim tawm hauv cov ntawv sau ua ke cov kauj ruam thiab tsim cov unigene teeb nrog isoform-theem daws teeb meem.Cov txheej txheem unigene no muab cov ntaub ntawv muaj txiaj ntsig zoo li "reference genome" ntawm transcriptome-theem.Tsis tas li ntawd, kev sib txuas nrog cov ntaub ntawv txuas ntxiv tom ntej, qhov kev pabcuam no txhawb nqa qhov tseeb ntawm isoform-theem qhia.
Platform: PacBio Sequel IILub tsev qiv ntawv: SMRT tswb tsev qiv ntawv -
Eukaryotic mRNA sequencing-Illumina
mRNA sequencing enables profiling ntawm tag nrho cov mRNAs transcripted los ntawm hlwb nyob rau hauv tej yam kev mob.Nws yog lub tshuab muaj zog rau kev nthuav tawm cov noob qhia profile, cov qauv noob thiab cov txheej txheem molecular ntawm qee cov txheej txheem lom neeg.Txog niaj hnub no, mRNA sequencing tau dav siv hauv kev tshawb fawb, kev kuaj mob, kev tsim tshuaj, thiab lwm yam.
Platform: Illumina NovaSeq 6000
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Tsis siv neeg raws li mRNA sequencing-Illumina
mRNA sequencing txais yuav cov txheej txheem txuas ntxiv tiam tom ntej (NGS) txhawm rau ntes tus xa xov RNA (mRNA) daim ntawv Eukaryote ntawm lub sijhawm tshwj xeeb uas qee qhov tshwj xeeb ua haujlwm.Cov ntawv sau ntev tshaj plaws tau raug hu ua 'Unigene' thiab siv los ua cov txheej txheem siv rau kev soj ntsuam tom ntej, uas yog ib qho txiaj ntsig zoo los kawm txog cov txheej txheem molecular thiab kev tswj hwm kev sib txuas ntawm hom yam tsis siv.
Tom qab transcriptome cov ntaub ntawv los ua ke thiab unigene functional annotation
(1) SNP tsom xam, SSR tsom xam, CDS kev kwv yees thiab cov qauv noob yuav raug ua ntej.
(2) Kev ua kom muaj nuj nqis ntawm unigene qhia hauv txhua tus qauv yuav ua.
(3) Sib txawv ntawm cov unigenes ntawm cov qauv (los yog pab pawg) yuav raug pom raws li unigene qhia
(4) Kev sib sau ua ke, cov lus piav qhia ua haujlwm thiab kev txheeb xyuas qhov sib txawv ntawm cov unigenes sib txawv yuav raug ua
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Ntev tsis-coding sequencing-Illumina
Ntev tsis-coding RNAs (lncRNAs) yog ib hom RNA molecules nrog ntev tshaj 200 nt, uas yog characterized los ntawm tsis tshua muaj coding peev xwm.LncRNA, uas yog tus tswv cuab tseem ceeb hauv cov tsis-coding RNAs, feem ntau pom muaj nyob rau hauv nucleus thiab ntshav.Kev txhim kho hauv kev sib txuas thev naus laus zis thiab bioinformtics ua rau muaj kev txheeb xyuas ntau yam tshiab lncRNAs thiab koom nrog cov haujlwm lom neeg.Cov ntaub ntawv pov thawj qhia tau hais tias lncRNA tau koom nrog hauv kev tswj hwm epigenetic, kev tswj hwm kev hloov pauv thiab kev tswj xyuas tom qab.
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Me RNA sequencing-Illumina
Me RNA yog hais txog ib chav kawm ntawm cov tsis-coding RNA molecules uas feem ntau tsawg dua 200nt nyob rau hauv ntev, suav nrog micro RNA (miRNA), me me cuam tshuam RNA (siRNA), thiab piwi-interacting RNA (piRNA).
MicroRNA (miRNA) yog ib chav kawm ntawm endogenous me RNA nrog ntev li ntawm 20-24nt, uas plays ntau yam tseem ceeb ntawm kev tswj lub luag haujlwm hauv hlwb.miRNA koom nrog ntau cov txheej txheem ntawm lub neej uas nthuav tawm cov ntaub so ntswg - tshwj xeeb thiab theem - kev qhia tshwj xeeb thiab tau txais txiaj ntsig zoo hauv ntau hom.
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CircRNA sequencing-Illumina
Tag nrho cov ntawv sau ua ke yog tsim los qhia txhua hom RNA molecules, suav nrog coding (mRNA) thiab tsis-coding RNAs (xws li lncRNA, circRNA thiab miRNA) uas tau sau los ntawm cov hlwb tshwj xeeb ntawm qee lub sijhawm.Tag nrho cov ntawv sau ua ke, tseem hu ua "tag nrho RNA sequencing" lub hom phiaj ntawm kev nthuav tawm cov kev tswj hwm kev sib txuas lus ntawm qib transcriptome.Ua kom zoo dua ntawm NGS thev naus laus zis, cov kab ke ntawm tag nrho cov khoom lag luam transcriptome muaj rau kev soj ntsuam ceRNA thiab kev sib koom ua ke RNA, uas muab thawj kauj ruam ntawm kev ua haujlwm zoo.Tshaj tawm kev tswj hwm network ntawm circRNA-miRNA-mRNA raws li ceRNA.
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Tag nrho cov ntawv sau ua ke - Illumina
Tag nrho cov ntawv sau ua ke yog tsim los qhia txhua hom RNA molecules, suav nrog coding (mRNA) thiab tsis-coding RNAs (xws li lncRNA, circRNA thiab miRNA) uas tau sau los ntawm cov hlwb tshwj xeeb ntawm qee lub sijhawm.Tag nrho cov ntawv sau ua ke, tseem hu ua "tag nrho RNA sequencing" lub hom phiaj ntawm kev nthuav tawm cov kev tswj hwm kev sib txuas lus ntawm qib transcriptome.Ua kom zoo dua ntawm NGS thev naus laus zis, cov kab ke ntawm tag nrho cov khoom lag luam transcriptome muaj rau kev soj ntsuam ceRNA thiab kev sib koom ua ke RNA, uas muab thawj kauj ruam ntawm kev ua haujlwm zoo.Tshaj tawm kev tswj hwm network ntawm circRNA-miRNA-mRNA raws li ceRNA.
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Prokaryotic RNA sequencing
Prokaryotic RNA sequencing siv cov tiam tom ntej sequencing (NGS) los qhia qhov muaj thiab ntau ntawm RNA ntawm ib lub sijhawm, los ntawm kev tshuaj xyuas qhov hloov pauv ntawm cov ntaub ntawv ntawm tes.Peb lub tuam txhab prokaryotic RNA sequencing, tshwj xeeb yog lub hom phiaj ntawm prokaryotes nrog kev siv genomes, muab koj cov ntaub ntawv sau tseg, kev txheeb xyuas cov qauv noob, thiab lwm yam.
Platform: Illumina NovaSeq 6000
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Metatranscriptome Sequencing
Metatranscriptome sequencing indentifies gene qhia ntawm microbes (ob leeg eukaryotes thiab prokaryotes) nyob rau hauv tej yam ntuj tso ib puag ncig (xws li av, dej, hiav txwv, quav, thiab plab.). ntawm hom, kev ua kom muaj txiaj ntsig zoo ntawm kev qhia cov noob sib txawv, thiab lwm yam.
Platform: Illumina NovaSeq 6000
