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Single-nucleus RNA Sequencing
Kutsogola kwa kujambula kwa selo limodzi ndi njira yopangira laibulale yamunthu payekha kuphatikiza ndi kutsata kopitilira muyeso kumathandizira maphunziro amtundu wa gene pa cell-by-cell.Imathandizira kusanthula kozama komanso kokwanira pama cell ovuta, momwe imapewa kubisala kusiyanasiyana kwawo potenga pafupifupi ma cell onse.
Komabe, ma cell ena sali oyenera kupangidwa kukhala kuyimitsidwa kwa selo limodzi, kotero njira zina zokonzekera zitsanzo - kuchotsa ma nucleus kuchokera ku minofu kumafunika, ndiko kuti, nyukiliya imachotsedwa mwachindunji ku minofu kapena selo ndikukonzedwa kuti ikhale imodzi-nucleus kuyimitsidwa kwa single- kutsatizana kwa ma cell.
BMK imapereka 10 × Genomics Chromium TM yochokera ku cell single RNA sequencing service.Utumikiwu wagwiritsidwa ntchito kwambiri pamaphunziro okhudzana ndi matenda, monga kusiyanitsa kwa maselo a chitetezo chamthupi, chotupa chosiyana, kukula kwa minofu, ndi zina zambiri.
Spatial transcriptome chip: 10 × Genomics
Pulatifomu: Illumina NovaSeq Platform
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BMKMANU S1000 Spatial Transcriptome
Spatial transcriptomics imatsogola pazatsopano zasayansi, kupatsa mphamvu ofufuza kuti afufuze movutikira momwe ma jini amafotokozera mkati mwa minofu ndikusunga malo awo.Pakati pa nsanja zosiyanasiyana, BMKGene yapanga BMKManu S1000 Spatial Transcriptome Chip, modzitamandira.chiganizo chowonjezerekaya 5µM, kufika pamtundu wa subcellular, ndikuthandiziramakonda amitundu yambiri.Chip cha S1000, chokhala ndi mawanga pafupifupi 2 miliyoni, chimagwiritsa ntchito tinthu tating'onoting'ono tomwe timakhala ndi mikanda yodzaza ndi zojambulidwa za barcode.Laibulale ya cDNA, yopangidwa ndi ma barcode a malo, imakonzedwa kuchokera ku chipangizo cha S1000 ndikutsatiridwa papulatifomu ya Illumina NovaSeq.Kuphatikiza kwa zitsanzo zokhala ndi barcode ndi UMIs zimatsimikizira kulondola ndi kutsimikizika kwa deta yopangidwa.Khalidwe lapadera la BMKManu S1000 chip lili mu kusinthasintha kwake, lopereka zosintha zamitundu ingapo zomwe zimatha kusinthidwa bwino kumitundu yosiyanasiyana komanso mwatsatanetsatane.Kusinthasintha kumeneku kumayika chip ngati chisankho chabwino kwambiri pamaphunziro osiyanasiyana amtundu wapamalo, kuwonetsetsa kusanja kolondola kwa malo popanda phokoso lochepa.
Pogwiritsa ntchito chipangizo cha BMKManu S1000 ndi matekinoloje ena a spatial transcriptomics, ofufuza atha kumvetsetsa bwino momwe ma cell amagwirira ntchito komanso kuyanjana kwa ma cell komwe kumachitika mkati mwa minyewa, ndikupereka chidziwitso chamtengo wapatali pamakina omwe amachokera kuzinthu zachilengedwe m'magawo osiyanasiyana, kuphatikiza. oncology, neuroscience, chitukuko cha biology, immunology ndi maphunziro a botanical.
Platform: BMKManu S1000 chip ndi Illumina NovaSeq
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10x Genomics Visium Spatial Transcriptome
Spatial transcriptomics ndiukadaulo wotsogola womwe umalola ofufuza kuti afufuze momwe ma jini amatchulidwira mkati mwa minofu ndikusunga malo awo.Pulatifomu imodzi yamphamvu mderali ndi 10x Genomics Visium yophatikizidwa ndi kutsatizana kwa Illumina.Mfundo ya 10X Visium yagona pa chip chapadera chokhala ndi malo ojambulidwa omwe zigawo za minofu zimayikidwa.Malo ogwidwawa ali ndi mabala a barcode, aliwonse ofanana ndi malo apadera mkati mwa minofu.Mamolekyu a RNA omwe adagwidwa kuchokera m'minyewayo amalembedwa ndi zozindikiritsa mamolekyulu (UMIs) panthawi yolemba mobwereza.Malo okhala ndi barcode awa ndi ma UMI amathandizira kupanga mapu olondola a malo komanso kuchuluka kwa mafotokozedwe a jini pakusintha kwa selo limodzi.Kuphatikiza kwa zitsanzo zokhala ndi barcode ndi UMIs zimatsimikizira kulondola ndi kutsimikizika kwa deta yopangidwa.Pogwiritsa ntchito ukadaulo uwu wa Spatial Transcriptomics, ofufuza amatha kumvetsetsa mozama momwe ma cell amakhalira komanso momwe ma cell amagwirira ntchito mkati mwa minyewa, ndikupereka chidziwitso chamtengo wapatali pamakina omwe amachokera kuzinthu zachilengedwe m'magawo angapo, kuphatikiza oncology, neuroscience, biology yachitukuko, chitetezo chamthupi. , ndi maphunziro a botanical.
Platform: 10X Genomics Visium ndi Illumina NovaSeq
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Kutalika Kwathunthu kwa mRNA Sequencing-Nanopore
Ngakhale kutsatizana kwa mRNA kochokera ku NGS kumagwira ntchito ngati chida chosunthika pakuchulukira kwa ma gene, kudalira kwake pamawerengedwe amfupi kumalepheretsa kugwira ntchito kwake pakuwunikira zovuta zolembera.Kutsata kwa Nanopore, kumbali ina, kumagwiritsa ntchito ukadaulo wowerengera nthawi yayitali, zomwe zimathandizira kutsatizana kwa zolembedwa zazitali za mRNA.Njirayi imathandizira kufufuza mwatsatanetsatane njira zina zophatikizira, kuphatikiza ma jini, poly-adenylation, komanso kuchuluka kwa ma isoforms a mRNA.
Kutsata kwa Nanopore kumadalira ma siginecha amagetsi a nanopore single-molecule nthawi yeniyeni.Motsogozedwa ndi mapuloteni amagalimoto, DNA yamitundu iwiri imamangiriza ku mapuloteni a nanopore ophatikizidwa mu biofilm, kumasuka pamene ikudutsa munjira ya nanopore pansi pa kusiyana kwamagetsi.Zizindikiro zamagetsi zapadera zomwe zimapangidwa ndi maziko osiyanasiyana pa chingwe cha DNA zimazindikirika ndikugawidwa mu nthawi yeniyeni, zomwe zimathandizira kutsata kolondola komanso kosalekeza kwa nucleotide.Njira yatsopanoyi imagonjetsa malire owerengeka pang'ono ndipo imapereka nsanja yosinthika yowunikira ma genomic, yomwe imaphatikizapo maphunziro ovuta a transcriptomic.
Platform: Nanopore Promethion P48
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Kutsata kutalika kwa mRNA -PacBio
Ngakhale kutsatizana kwa mRNA kochokera ku NGS ndi chida chogwiritsa ntchito mosiyanasiyana pakuwerengera ma jini, kudalira kwake pamawerengedwe afupikitsa kumalepheretsa kugwiritsidwa ntchito kwake pakuwunikira zovuta zolembera.PacBio sequencing (Iso-Seq), kumbali ina, imagwiritsa ntchito ukadaulo wowerengera nthawi yayitali, zomwe zimathandizira kutsatizana kwa zolembedwa zazitali za mRNA.Njirayi imathandizira kufufuza mozama kwa njira zina zophatikizira, kuphatikizika kwa jini ndi poly-adenylation ngakhale sichosankha choyambirira cha gene expression quantification, chifukwa cha kuchuluka kwa data yomwe ikufunika.
Ukadaulo wotsatizana wa PacBio umadalira kutsatizana kwa molekyulu imodzi, nthawi yeniyeni (SMRT), kupereka mwayi wapadera wojambula zolemba zazitali za mRNA.Njira yatsopanoyi imaphatikizapo kugwiritsa ntchito ma zero-mode waveguides (ZMWs), zitsime zokhala ndi microfabricated zomwe zimathandiza kuti nthawi yeniyeni ya DNA polymerase iwonetsedwe panthawi yotsatizana.Mkati mwa ma ZMW awa, PacBio's DNA polymerase imapanga ulusi wowonjezera wa DNA, ndikupanga zowerengera zazitali zomwe zimatenga zolemba zonse za mRNA.Opaleshoni ya PacBio mumayendedwe a Circular Consensus sequencing (CCS) imakulitsa kulondola mwa kutsata mobwerezabwereza molekyulu yomweyo.Mawerengedwe a HiFi omwe amapangidwa ali ndi kulondola kofanana ndi NGS, zomwe zimathandizira pakuwunika kokwanira komanso kodalirika kwa zovuta zolembera.Platform: PacBio Sequel II
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Eukaryotic mRNA Sequencing-Illumina
Kutsatizana kwa mRNA kumapatsa mphamvu kusanthula kwatsatanetsatane kwa zolembedwa zonse za mRNA m'maselo malinga ndi momwe zinthu ziliri.Ukadaulo wotsogolawu umagwira ntchito ngati chida champhamvu, kuwulula mbiri yakale ya jini, kapangidwe ka majini, ndi njira zama cell zomwe zimagwirizanitsidwa ndi mitundu yosiyanasiyana yachilengedwe.Kutengera kwambiri pakufufuza kofunikira, zowunikira zamankhwala, komanso kakulidwe ka mankhwala, kutsatizana kwa mRNA kumapereka chidziwitso pazovuta zama cellular dynamics ndi genetic regulation.
Platform: Illumina NovaSeq X
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Non-Reference based mRNA Sequencing-Illumina
Kutsatizana kwa mRNA kumapatsa mphamvu kusanthula kwatsatanetsatane kwa zolembedwa zonse za mRNA m'maselo malinga ndi momwe zinthu ziliri.Ukadaulo wotsogolawu umagwira ntchito ngati chida champhamvu, kuwulula mbiri yakale ya jini, kapangidwe ka majini, ndi njira zama cell zomwe zimagwirizanitsidwa ndi mitundu yosiyanasiyana yachilengedwe.Kutengera kwambiri pakufufuza kofunikira, zowunikira zamankhwala, komanso kakulidwe ka mankhwala, kutsatizana kwa mRNA kumapereka chidziwitso pazovuta zama cellular dynamics ndi genetic regulation.
Platform: Illumina NovaSeq X
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Kutsata kwanthawi yayitali kosalemba-Illumina
Ma RNA (lncRNA) autali osalemba ma RNA ndi ma RNA otalika kuposa ma nucleotide 200 omwe ali ndi kuthekera kocheperako ndipo ndi zinthu zofunika kwambiri mkati mwa RNA yosalemba.Zopezeka mu nucleus ndi cytoplasm, ma RNA awa amatenga gawo lofunikira pakuwongolera epigenetic, transcriptal, ndi post-transcriptal, kutsimikizira kufunikira kwawo pakupanga ma cell ndi ma cell.Kutsata kwa LncRNA ndi chida champhamvu pakusiyanitsa ma cell, Ontogenesis, ndi matenda a anthu.
Platform: Illumina NovaSeq
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Small RNA sequencing-Illumina
Mamolekyu ang'onoang'ono a RNA (sRNA), omwe nthawi zambiri amakhala pansi pa 200 nucleotide m'litali, amaphatikizapo ma microRNAs (miRNAs), ma RNAs (siRNAs), ndi ma RNA (piRNAs) omwe amasokoneza piwi.Mwa awa, ma miRNA, ozungulira ma nucleotide 20-24 aatali, ndiwodziwika kwambiri chifukwa cha maudindo awo ofunikira pamachitidwe osiyanasiyana am'manja.Pokhala ndi mawonekedwe okhudzana ndi minofu komanso siteji, ma miRNA amawonetsa kusamala kwambiri pamitundu yosiyanasiyana.
Platform: Illumina NovaSeq
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circRNA sequencing-Illumina
Zozungulira RNA sequencing (circRNA-seq) ndikuwonetsa ndi kusanthula ma RNA ozungulira, gulu la mamolekyu a RNA omwe amapanga malupu otsekedwa chifukwa cha zochitika zomwe sizinali za canonical splicing, zomwe zimapatsa RNA iyi kukhala yokhazikika.Ngakhale ma circRNA ena awonetsedwa kuti amachita ngati masiponji a microRNA, kuthamangitsa ma microRNA ndikuwalepheretsa kuwongolera ma mRNAs awo, ma circRNA ena amatha kulumikizana ndi mapuloteni, kusintha mawonekedwe a jini, kapena kukhala ndi maudindo pama cell.Kusanthula kwa mawu a circRNA kumapereka chidziwitso pamaudindo owongolera mamolekyuwa komanso kufunikira kwawo munjira zosiyanasiyana zama cell, magawo akukula, ndi matenda, zomwe zimathandizira kumvetsetsa mozama za zovuta zamalamulo a RNA potengera mafotokozedwe amtundu.
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Kutsatizana konse kwa transcriptome - Illumina
Kutsatizana kwa zolemba zonse kumapereka njira yokwanira yolembera mamolekyu osiyanasiyana a RNA, kuphatikiza ma RNA onse (mRNA) ndi ma RNA osalemba (lncRNA, circRNA, ndi miRNA).Njirayi imagwira zolemba zonse zama cell enieni panthawi inayake, kulola kumvetsetsa kwathunthu kwa ma cell.Zomwe zimatchedwanso "total RNA sequencing," cholinga chake ndi kuwulula maukonde owongolera pamlingo wa transcriptome, kupangitsa kusanthula mozama monga kupikisana kwa RNA (ceRNA) ndi kusanthula kwa RNA.Izi ndizomwe zimayambira poyambira kuzindikirika kwa magwiridwe antchito, makamaka pakuwulula maukonde owongolera omwe amakhudza kuyanjana kwa ceRNA kochokera ku circRNA-miRNA-mRNA.
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Kutsata kwa Prokaryotic mRNA
Kutsatizana kwa mRNA kumapatsa mphamvu kusanthula kwatsatanetsatane kwa zolembedwa zonse za mRNA m'maselo malinga ndi momwe zinthu ziliri.Ukadaulo wotsogolawu umagwira ntchito ngati chida champhamvu, kuwulula mbiri yakale ya jini, kapangidwe ka majini, ndi njira zama cell zomwe zimagwirizanitsidwa ndi mitundu yosiyanasiyana yachilengedwe.Kutengera kwambiri pakufufuza kofunikira, zowunikira zamankhwala, komanso kakulidwe ka mankhwala, kutsatizana kwa mRNA kumapereka chidziwitso pazovuta zama cellular dynamics ndi genetic regulation.Kukonzekera kwathu kwachitsanzo kwa prokaryotic mRNA kumapangidwira zolemba za prokaryotic, zomwe zikuphatikiza kuchepa kwa rRNA komanso kukonzekera kwama library.
Platform: Illumina NovaSeq X
