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I-nucleus ye-RNA enye
Ukuqhubela phambili ekuthathweni kweseli enye kunye nobuchule bokwakha ithala leencwadi ngalinye kudityaniswe nolandelelwano olugqithileyo luvumela izifundo zokuchazwa kofuzo ngokwesiseko seseli-nge-seli.Ivumela uhlalutyo olunzulu kunye olupheleleyo lwenkqubo kwiiseli ezintsonkothileyo, apho ubukhulu becala inqanda ukufihla ukungafani kwazo ngokuthatha umndilili wazo zonke iiseli.
Nangona kunjalo, ezinye iiseli azifanelekanga ukuba zenziwe zibe yi-single-cell suspension, ngoko ke ezinye iindlela zokulungiselela iisampulu-i-nucleus extraction evela kwizicubu ziyafuneka, oko kukuthi, i-nucleus ikhutshwa ngokuthe ngqo kwiithishu okanye kwiseli kwaye ilungiswe ibe yi-single-nucleus suspension for single- ulandelelwano lweseli.
I-BMK inikezela nge-10 × i-Genomics yeChromium TM esekelwe kwinkonzo yokulandelelana kwe-RNA yeseli enye.Le nkonzo isetyenziswe ngokubanzi kwizifundo kwizifundo ezinxulumene nesifo, ezifana nokwahlukana kweeseli zomzimba, i-tumor heterogeneity, uphuhliso lwezicubu, njl.
I-Spatial transcriptome chip: 10× Genomics
Iqonga: I-Illumina NovaSeq Platform
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Ulandelelwano lweGenome yeSityalo/yeSilwanyana
Ukulandelelana kwakhona kwe-genome yonke, eyaziwa ngokuba yi-WGS, yenza ukuba kutyhilwe zombini iinguqulelo eziqhelekileyo kunye nezinqabileyo kuyo yonke i-genome kuquka i-Single Nucleotide Polymorphism (SNP), ukucinywa kokuFaka (InDel), ukwahluka koLwakhiwo (SV), kunye neCopy Number Variation (CNV). ).I-SVs zenza inxalenye enkulu yesiseko sokwahluka kune-SNP kwaye ibe nempembelelo enkulu kwi-genome, enempembelelo ebalulekileyo kwizinto eziphilayo.Ukuphinda kufundwe ixesha elide kuvumela ukuchongwa okuchane ngakumbi kwamaqhekeza amakhulu kunye nokwahluka okuntsokothileyo kuba ukufunda ixesha elide kwenza kube lula ukuwela ichromosomal ngaphaya kwemimandla entsonkothileyo efana nokuphindaphinda kwe-tandem, imimandla etyebileyo yeGC/AT, kunye nemimandla eguquguqukayo.
Iqonga: Illumina, PacBio, Nanopore
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I-BMKMANU S1000 i-Spatial Transcriptome
I-Spatial transcriptomics imi phambili kubuchule obutsha bezenzululwazi, ixhobisa abaphandi ukuba baphengulule kwiipateni ezintsonkothileyo zokubonakaliswa kofuzo ngaphakathi kwezicubu ngelixa begcina imeko yabo yendawo.Phakathi kwamaqonga ahlukeneyo, iBMKGene iphuhlise iBMKManu S1000 Spatial Transcriptome Chip, iqhayisaisisombululo esongeziweyoye 5µM, ifikelela kuluhlu olusezantsi, kunye nokwenzauseto lwesisombululo esinamanqanaba amaninzi.Itshiphu ye-S1000, enamabala amalunga nezigidi ezi-2, isebenzisa ii-microwells ezalekwe ngamaso alayishwe nge-spatially barcoded capture probes.Ilayibrari ye-cDNA, etyetyiswe ngeebhakhowudi zendawo, ilungiswa ukusuka kwitshiphu ye-S1000 kwaye emva koko ilandelelaniswe kwiqonga le-Illumina NovaSeq.Ukudityaniswa kweesampulu ezinebhakhowudi yendawo kunye nee-UMIs ziqinisekisa ukuchaneka kunye nokuchaneka kwedatha eyenziweyo.Uphawu olukhethekileyo lwe-chip ye-BMKManu S1000 ilele kukuguquguquka kwayo, inika useto lwesisombululo esinamanqanaba amaninzi anokuthi alungelelaniswe kakuhle kwiithishu ezahlukeneyo kunye namanqanaba eenkcukacha.Oku kuguquguquka kubeka itshiphu njengokhetho olubalaseleyo lwezifundo ezahlukeneyo zoshicilelo lwesithuba, ukuqinisekisa ukudityaniswa kwendawo echanekileyo kunye nengxolo encinci.
Ukusebenzisa i-chip ye-BMKManu S1000 kunye nobunye ubuchwephesha be-transcriptomics yendawo, abaphandi banokufumana ukuqonda okungcono kwentlangano yendawo yeeseli kunye nokunxibelelana kweemolekyuli ezintsonkothileyo ezenzeka ngaphakathi kwezicubu, zibonelela ngolwazi oluxabisekileyo kwiinkqubo eziphantsi kwenkqubo yebhayoloji kuluhlu olubanzi lweenkalo, kubandakanya. i-oncology, i-neuroscience, i-biology yophuhliso, i-immunology kunye nezifundo zebhotani.
Iqonga: BMKManu S1000 chip kunye Illumina NovaSeq
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I-10x ye-Genomics yeVisium ye-Spatial Transcriptome
I-Spatial transcriptomics yitekhnoloji enqamlezileyo evumela abaphandi ukuba baphande iipatheni zokubonakaliswa kofuzo ngaphakathi kwezicubu ngelixa begcina imeko yabo yendawo.Iqonga elinye elinamandla kule domain yi-10x Genomics Visium edityaniswa nokulandelelana kwe-Illumina.Umgaqo we-10X Visium ulele kwi-chip ekhethekileyo enendawo ekhethiweyo yokubamba apho amacandelo e-tissue abekwe khona.Le ndawo yokubamba iqulethe iindawo ezinebhakhowudi, nganye ihambelana nendawo eyodwa ngaphakathi kwethishu.Iimolekyuli ze-RNA ezibanjiweyo kwiithishu zibhalwa emva koko zibhalwe nge-molecular identifiers (UMIs) ezikhethekileyo ngexesha lenkqubo yokubhala umva.Ezi ndawo zinebhakhowudi kunye nee-UMIs zenza imephu yendawo echanekileyo kunye nobungakanani bokubonakaliswa kofuzo kwisisombululo seseli enye.Ukudityaniswa kweesampulu ezinebhakhowudi yendawo kunye nee-UMIs ziqinisekisa ukuchaneka kunye nokuchaneka kwedatha eyenziweyo.Ngokusebenzisa le teknoloji ye-Spatial Transcriptomics, abaphandi banokufumana ukuqonda okunzulu malunga nentlangano yendawo yeeseli kunye nentsebenziswano eyinkimbinkimbi ye-molecular eyenzeka ngaphakathi kwezicubu, inika ulwazi oluxabisekileyo kwiindlela eziphantsi kweenkqubo zebhayoloji kwiinkalo ezininzi, kuquka i-oncology, neuroscience, i-biology yophuhliso, i-immunology. , kunye nezifundo zebhotani.
Iqonga: I-10X Genomics Visium kunye ne-Illumina NovaSeq
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Ubude obupheleleyo be-mRNA Ukulandelelana-Nanopore
Ngelixa ulandelelwano lwe-mRNA olusekwe kwi-NGS lusebenza njengesixhobo esiguquguqukayo sobungakanani bokuchazwa kofuzo, ukuthembela kufundo olufutshane kuthintela ukusebenza kwayo kuhlalutyo oluntsonkothileyo lwe-transcriptomic.Ulandelelwano lweNanopore, kwelinye icala, lusebenzisa itekhnoloji yokufunda ixesha elide, eyenza ulandelelwano lwemibhalo ebhaliweyo ye-mRNA yobude obupheleleyo.Le ndlela iququzelela ukuphononongwa okubanzi kwezinye iindlela zokudityaniswa, ukudityaniswa kwemfuza, i-poly-adenylation, kunye nobungakanani be-mRNA isoforms.
Ulandelelwano lweNanopore luxhomekeke kwi-nanopore molekyuli enye yexesha lokwenyani lombane.Ukukhokelwa ziiprotheyini zeemoto, i-DNA ephindwe kabini ibophelela kwiiprotheni ze-nanopore ezifakwe kwi-biofilm, ihlanjululwe njengoko idlula kumjelo we-nanopore phantsi kokwahlukana kombane.Izibonakaliso zombane ezihlukeneyo eziveliswa ziziseko ezahlukeneyo kwi-DNA strand zichongwa kwaye zihlelwe ngexesha langempela, ziququzelele ukulandelelana kwe-nucleotide echanekileyo kunye nokuqhubekayo.Le ndlela intsha yoyisa imida yokufunda emfutshane kwaye ibonelela ngeqonga eliguquguqukayo lohlalutyo oluntsonkothileyo lwe-genomic, lubandakanya izifundo ezintsonkothileyo ze-transcriptomic.
Iqonga: iNanopore Promethion P48
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Ulandelelwano olupheleleyo lwe-mRNA -PacBio
Ngelixa ulandelelwano olusekwe kwi-NGS ye-mRNA sisixhobo esiguquguqukayo sokulinganisa ukubonakaliswa kofuzo, ukuthembela kufundo olufutshane kuthintela ukusetyenziswa kwayo kuhlalutyo oluntsonkothileyo lwe-transcriptomic.Ukulandelelana kwe-PacBio (i-Iso-Seq), ngakolunye uhlangothi, isebenzisa iteknoloji yokufunda ixesha elide, eyenza ukulandelelana kwemibhalo egcweleyo ye-mRNA.Le ndlela iququzelela uphononongo olubanzi lwezinye iindlela zokudityaniswa, ukudityaniswa kwemfuza kunye ne-poly-adenylation nangona ingeyiyo eyona nto iphambili yokukhetha ubungakanani bokuchazwa kofuzo, ngenxa yedatha ephezulu efunekayo.
Itekhnoloji yokulandelelanisa i-PacBio ixhomekeke kwi-molecule enye, i-real-time (SMRT) yokulandelelana, inika inzuzo eyahlukileyo ekubanjeni imibhalo ye-mRNA yobude obupheleleyo.Le ndlela entsha ibandakanya ukusetyenziswa kwe-zero-mode waveguides (ZMWs), imingxuma emincinci eyenza ukubonwa kwexesha langempela lomsebenzi we-DNA polymerase ngexesha lokulandelelana.Ngaphakathi kwezi ZMWs, iPacBio's DNA polymerase idibanisa umsonto ohambelanayo weDNA, ivelisa ufundo olude olufikelela kuyo yonke imibhalo ye-mRNA.Ukusebenza kwe-PacBio kwimodi yeSetyhula ye-Consensus sequencing (CCS) iphucula ukuchaneka ngokulandelelana ngokuphindaphindiweyo i-molecule efanayo.Ukufundwa kwe-HiFi eveliswayo kunokuchaneka okuthelekiswa ne-NGS, igalelo elingakumbi kuhlalutyo olubanzi noluthembekileyo lweempawu ezintsonkothileyo ze-transcriptomic.Iqonga: I-PacBio Sequel II
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Eukaryotic mRNA Sequencing-Illumina
Ulandelelwano lwe-mRNA luxhobisa iprofayili ebanzi yazo zonke iikopi ze-mRNA ngaphakathi kweeseli phantsi kweemeko ezithile.Le teknoloji ye-cutting-edge isebenza njengesixhobo esinamandla, ityhila iiprofayili zegenesis ezintsonkothileyo, ulwakhiwo lwemfuza, kunye neendlela zemolekyuli ezinxulumene neenkqubo ezahlukeneyo zebhayoloji.Yamkelwa ngokubanzi kuphando olusisiseko, uxilongo lweklinikhi, kunye nophuhliso lwamachiza, ulandelelwano lwe-mRNA lubonelela ngokuqonda kubuntsonkotha bokutshintshatshintsha kweeseli kunye nolawulo lwemfuzo.
Iqonga: Illumina NovaSeq X
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Non-Reference esekelwe mRNA Sequencing-Illumina
Ulandelelwano lwe-mRNA luxhobisa iprofayili ebanzi yazo zonke iikopi ze-mRNA ngaphakathi kweeseli phantsi kweemeko ezithile.Le teknoloji ye-cutting-edge isebenza njengesixhobo esinamandla, ityhila iiprofayili zegenesis ezintsonkothileyo, ulwakhiwo lwemfuza, kunye neendlela zemolekyuli ezinxulumene neenkqubo ezahlukeneyo zebhayoloji.Yamkelwa ngokubanzi kuphando olusisiseko, uxilongo lweklinikhi, kunye nophuhliso lwamachiza, ulandelelwano lwe-mRNA lubonelela ngokuqonda kubuntsonkotha bokutshintshatshintsha kweeseli kunye nolawulo lwemfuzo.
Iqonga: Illumina NovaSeq X
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Ixesha elide non-coding ulandelelwano-Illumina
Ii-RNA ezinde ezingakhowudiyo (lncRNAs) zii-RNAs ezinde kune-200 nucleotides ezinomthamo omncinci wokubhala iikhowudi kwaye zizinto ezibalulekileyo ngaphakathi kwe-RNA engakhowudi.Ifunyanwe kwi-nucleus kunye ne-cytoplasm, ezi RNAs zidlala indima ebalulekileyo kwi-epigenetic, transcriptal, kunye ne-post-transcriptal regulation, igxininisa ukubaluleka kwazo ekubumbeni iinkqubo zeselula kunye ne-molecular.Ukulandelelana kwe-LncRNA sisixhobo esinamandla kwi-Cell differentiation, i-Ontogenesis, kunye nezifo zabantu.
Iqonga: Illumina NovaSeq
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I-RNA encinci yokulandelelana-Illumina
Iimolekyuli ezincinci ze-RNA (sRNA), ngokuqhelekileyo phantsi kwe-nucleotides ye-200 ubude, zibandakanya i-microRNAs (miRNAs), i-RNAs encinci ephazamisayo (siRNAs), kunye ne-piwi-interacting RNAs (piRNAs).Phakathi kwezi, ii-miRNAs, malunga ne-20-24 nucleotides ubude, ziphawuleka ngakumbi kwiindima zazo zolawulo olubalulekileyo kwiinkqubo ezahlukeneyo zeselula.Ngeepateni zokuchaza ezithe ngqo kunye nenqanaba elithile, ii-miRNAs zibonisa ulondolozo oluphezulu kuzo zonke iintlobo ezahlukeneyo.
Iqonga: Illumina NovaSeq
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circRNA ulandelelwano-Illumina
Ulandelelwano lwe-RNA yeSetyhula (i-circRNA-seq) kukuchaza iphrofayili kunye nokuhlalutya i-RNAs ejikelezayo, iklasi yee-molecule ze-RNA ezenza i-loops evaliweyo ngenxa yeziganeko ze-non-canonical splicing, ezibonelela ezi RNA ngokunyuka kozinzo.Ngelixa ezinye ii-circRNAs zibonakaliswe ukuba zisebenza njengeziponji ze-microRNA, zihlutha ii-microRNAs kwaye zibathintele ekulawuleni i-mRNAs ekujoliswe kuyo, ezinye ii-circRNAs zinokusebenzisana neeprotheni, zimodareyitha imbonakalo yemfuza, okanye zibe neendima kwiinkqubo zeselula.Uhlalutyo lwentetho ye-circRNA lubonelela ngengqiqo kwiindima zokulawula ezi molekyuli kunye nokubaluleka kwazo kwiinkqubo ezahlukeneyo zeselula, amanqanaba ophuhliso, kunye neemeko zezifo, ezinegalelo ekuqondeni okunzulu kobunzima bolawulo lwe-RNA kumxholo we-gene expression.
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Ulandelelwano olupheleleyo lwe-transcriptome - Illumina
Ulandelelwano olupheleleyo lwe-transcriptome lubonelela ngendlela ebanzi yokuchaza iimolekyuli ezahlukeneyo ze-RNA, eziquka zombini iikhowudi (mRNA) kunye nee-RNA ezingakhowudi (lncRNA, circRNA, kunye ne-miRNA).Obu buchule bubamba i-transcriptome epheleleyo yeeseli ezithile ngexesha elithile, ivumela ukuqonda okupheleleyo kweenkqubo zeselula.Eyaziwa nangokuthi "ulandelelwano lwe-RNA iyonke," ijolise ekutyhileni uthungelwano lolawulo oluntsonkothileyo kwinqanaba le-transcriptome, ivumela uhlalutyo olunzulu olufana ne-RNA ekhuphisanayo ye-endogenous (ceRNA) kunye nohlalutyo lwe-RNA edibeneyo.Oku kuphawula inyathelo lokuqala elibhekiselele kuphawu olusebenzayo, ngakumbi ekutyhileni uthungelwano lolawulo olubandakanya i-circRNA-miRNA-mRNA-esekwe kunxibelelwano lwe-ceRNA.
