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Ubude obugcweleyo bolandelelwano lwe-mRNA-Nanopore
Ulandelelwano lwe-RNA ibe sisixhobo esixabisekileyo sohlalutyo olupheleleyo lwe-transcriptome.Ngokungathandabuzekiyo, ulandelelwano lokufunda okufutshane lwemveli luzuze uphuhliso oluninzi olubalulekileyo apha.Nangona kunjalo, ihlala idibana nemida ekuchongeni kwe-isoform yobude obugcweleyo, ubungakanani, ukuthambekela kwePCR.
Ukulandelelana kwe-Nanopore kuyahlula kwezinye iiplatifomu zokulandelelana, kuba i-nucleotides ifundwa ngokuthe ngqo ngaphandle kwe-DNA synthesis kwaye ivelise ukufunda ixesha elide kwishumi leekilobases.Oku kuxhobisa ukufundwa ngokuthe ngqo ukuwela imibhalo yobude obugcweleyo kunye nokujongana nemingeni kwizifundo zenqanaba le-isoform.
Iqonga:Nanopore PromethION
Ithala leencwadi:cDNA-PCR
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De novo Ubude obugcweleyo bokulandelelana kweTranscriptome -PacBio
De novoUlandelelwano olupheleleyo lwe-transcriptome, ekwaziwa njengeDe novoI-Iso-Seq ithatha iingenelo ze-PacBio sequencer kubude bokufunda, eyenza ukulandelelana kwee-molecule ze-cDNA ezigcweleyo ngaphandle kokuphazamiseka.Oku kuthintela ngokupheleleyo naziphi na iimpazamo eziveliswe kumanyathelo endibano yokubhala kwaye kwakha iiseti ze-unigene ezinesisombululo senqanaba le-isoform.Ezi seti ze-unigene zibonelela ngolwazi olunamandla lwemfuza njenge "reference genome" kwinqanaba le-transcriptome.Ukongeza, ukudibanisa nedatha yolandelelwano lwesizukulwana esilandelayo, le nkonzo ixhobisa ubungakanani obuchanekileyo be-isoform-level expression.
Iqonga: I-PacBio Sequel IIIthala leencwadi: Ilayibrari yentsimbi ye-SMRT -
Ulandelelwano lwe-Eukaryotic mRNA-Illumina
Ulandelelwano lwe-mRNA lwenza ukuba iprofayili yazo zonke i-mRNA zikhutshelwe kwiiseli phantsi kweemeko ezithile.Bubuchwephesha obunamandla bokutyhila inkangeleko yemfuza yemfuza, izakhiwo zemfuza kunye neendlela zemolekyuli zeenkqubo ezithile zebhayoloji.Ukuza kuthi ga ngoku, ulandelelwano lwe-mRNA lusetyenziswe ngokubanzi kuphando olusisiseko, uxilongo lwezonyango, uphuhliso lwamachiza, njl.
Iqonga: Illumina NovaSeq 6000
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Non-Reference esekelwe mRNA ulandelelwano-Illumina
Ulandelelwano lwe-mRNA lusebenzisa ubuchule bolandelelwano lwesizukulwana esilandelayo (NGS) ukubamba umthunywa i-RNA(mRNA) ifom ye-Eukaryote ngexesha elithile lemisebenzi ekhethekileyo eyenziwayo.Owona mbhalo mde uceliweyo wawubizwa ngokuba 'yi-Unigene' kwaye isetyenziswe njengolandelelwano lwereferensi kuhlalutyo olulandelayo, oluyindlela esebenzayo yokufunda umatshini wemolekyuli kunye nothungelwano lolawulo lweentlobo ngaphandle kwereferensi.
Emva kokuhlanganiswa kwedatha ye-transcriptome kunye nochazo olusebenzayo lwe-unigene
(1) Uhlalutyo lwe-SNP, uhlalutyo lwe-SSR, ukubikezelwa kwe-CDS kunye nesakhiwo sofuzo siya kwenziwa kwangaphambili.
(2) Ubungakanani bentetho ye-unigene kwisampulu nganye iya kwenziwa.
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(4) Ukuhlanganisana, uchazo olusebenzayo kunye nohlalutyo lokutyebisa lwee-unigenes ezichazwe ngokwahlukileyo ziya kwenziwa.
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Ixesha elide non-coding ulandelelwano-Illumina
Ii-RNAs ezinde ezingezizo iikhowudi (lncRNAs) luhlobo lweeamolekyuli ze-RNA ezinobude obungaphezulu kwe-200 nt, eziphawulwa ngokubanakho ukukhowuda okuphantsi kakhulu.I-LncRNA, njengelungu eliphambili kwii-RNA ezingakhowudi, ifumaneka ikakhulu kwi-nucleus kunye ne-plasma.Uphuhliso lolandelelwano lwetekhnoloji kunye ne-bioinformatics yenza ukuba kuchongwe iinoveli ezininzi ze-lncRNAs kwaye zinxulumanise ezo zinemisebenzi yebhayoloji.Ubungqina obuninzi bubonisa ukuba i-lncRNA ibandakanyeka ngokubanzi kulawulo lwe-epigenetic, umgaqo wokukhutshelwa kunye nommiselo emva kokubhala.
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I-RNA encinci yokulandelelana-Illumina
I-RNA encinci ibhekisela kwiklasi yee-molecule ze-RNA ezingenayo ikhowudi ezihlala zingaphantsi kwe-200nt ubude, kuquka i-RNA encinci (i-miRNA), i-RNA encinci (i-siRNA), kunye ne-piwi-interacting RNA (piRNA).
I-MicroRNA (miRNA) yiklasi ye-RNA encinci engapheliyo kunye nobude malunga ne-20-24nt, edlala indima ebalulekileyo yokulawula kwiiseli.I-miRNA ibandakanyeka kwiinkqubo ezininzi zobomi ezibonisa izicubu-ezithile kunye nenqanaba - ukubonakaliswa okuthe ngqo kunye nokulondolozwa kakhulu kwiintlobo ezahlukeneyo.
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CircRNA ulandelelwano-Illumina
Ulandelelwano olupheleleyo lwe-transcriptome luyilelwe ukwenza iprofayili yazo zonke iintlobo ze-molecule ze-RNA, kubandakanywa ikhowudi (mRNA) kunye nee-RNA ezingakhowudi (kuquka i-lncRNA, i-circRNA kunye ne-miRNA) ezibhalwe ngeeseli ezithile ngexesha elithile.Ulandelelwano lwe-transcriptome epheleleyo, ekwabizwa ngokuba "yitotali yolandelelwano lwe-RNA" ijolise ekuvezeni uthungelwano olubanzi lolawulo kwinqanaba le-transcriptome.Ukuthatha inzuzo yetekhnoloji ye-NGS, ulandelelwano lweemveliso ezipheleleyo ze-transcriptome ziyafumaneka kuhlalutyo lwe-ceRNA kunye nohlalutyo oludibeneyo lwe-RNA, olubonelela ngenyathelo lokuqala ekubonakalisweni kokusebenza.Ukutyhila inethiwekhi yolawulo ye-circRNA-miRNA-mRNA esekwe kwi-ceRNA.
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Ulandelelwano olupheleleyo lwe-transcriptome - Illumina
Ulandelelwano olupheleleyo lwe-transcriptome luyilelwe ukwenza iprofayili yazo zonke iintlobo ze-molecule ze-RNA, kubandakanywa ikhowudi (mRNA) kunye nee-RNA ezingakhowudi (kuquka i-lncRNA, i-circRNA kunye ne-miRNA) ezibhalwe ngeeseli ezithile ngexesha elithile.Ulandelelwano lwe-transcriptome epheleleyo, ekwabizwa ngokuba "yitotali yolandelelwano lwe-RNA" ijolise ekuvezeni uthungelwano olubanzi lolawulo kwinqanaba le-transcriptome.Ukuthatha inzuzo yetekhnoloji ye-NGS, ulandelelwano lweemveliso ezipheleleyo ze-transcriptome ziyafumaneka kuhlalutyo lwe-ceRNA kunye nohlalutyo oludibeneyo lwe-RNA, olubonelela ngenyathelo lokuqala ekubonakalisweni kokusebenza.Ukutyhila inethiwekhi yolawulo ye-circRNA-miRNA-mRNA esekwe kwi-ceRNA.
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Prokaryotic RNA ulandelelwano
Ukulandelelana kwe-Prokaryotic RNA kusebenzisa isizukulwana esilandelayo sokulandelelana (NGS) ukuveza ubukho kunye nobuninzi be-RNA ngexesha elithile, ngokuhlalutya ukuguqulwa kwe-transcriptome yeselula.Ulandelelwano lwe-RNA ye-prokaryotic yenkampani yethu, ijolise ngokukodwa kwiiprokaryotes ezinereferensi ye-genomes, ikubonelela ngeprofayili ye-transcriptome, uhlalutyo lwesakhiwo sofuzo, njl njl.
Iqonga: Illumina NovaSeq 6000
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Ulandelelwano lweMetatranscriptome
Ulandelelwano lweMetatranscriptome luchonga ukubonakaliswa kofuzo lwemicrobes (zombini ieukaryotes kunye neprokaryotes) ngaphakathi kokusingqongileyo kwendalo (oko kukuthi, umhlaba, amanzi, ulwandle, ilindle, kunye namathumbu.) Ngokukodwa, ezi nkonzo zikuvumela ukuba ufumane iprofayili yegenesis epheleleyo yentsholongwane entsonkothileyo yoluntu, uhlalutyo lwetaxonomic kweentlobo, uhlalutyo lokutyebisa olusebenzayo lwemfuza echazwe ngokwahlukileyo, kunye nokunye.
Iqonga: Illumina NovaSeq 6000
