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Hoʻokahi-nucleus RNA Sequencing
ʻO ka holomua o ka hopu ʻana i ke kelepona hoʻokahi a me ka ʻenehana kūkulu hale waihona puke me ka hui pū ʻana me ke kaʻina hana kiʻekiʻe e hiki ai i nā haʻawina hōʻike gene ma ke kumu cell-by-cell.Hāʻawi ia i kahi ʻōnaehana hohonu a piha hoʻi i nā heluna cell paʻakikī, kahi e pale nui ai i ka huna ʻana i ko lākou heterogeneity ma ka lawe ʻana i ka awelika o nā cell āpau.
Eia nō naʻe, ʻaʻole kūpono kekahi mau cell e hana ʻia i hoʻokuʻu ʻia hoʻokahi-cell, no laila, pono nā ʻano hana hoʻomākaukau ʻē aʻe - ka unuhi ʻana o ka nucleus mai nā ʻiʻo, ʻo ia hoʻi, lawe pololei ʻia ka nucleus mai nā ʻiʻo a i ʻole cell a hoʻomākaukau ʻia i hoʻokahi-nucleus suspension no hoʻokahi- ke kaʻina ʻana o ke kelepona.
Hāʻawi ʻo BMK i ka 10× Genomics ChromiumTM i hoʻokumu ʻia i ka lawelawe hoʻokaʻina RNA pū hoʻokahi.Ua hoʻohana nui ʻia kēia lawelawe i nā haʻawina e pili ana i nā haʻawina e pili ana i nā maʻi, e like me ka hoʻokaʻawale ʻana o ka cell immune, heterogeneity tumor, tissue development, etc.
Kīpī transcriptome āpau: 10× Genomics
Papahana: Illumina NovaSeq Platform
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Mea kanu/holoholona holoʻokoʻa Genome Sequencing
ʻO ka hoʻonohonoho hou ʻana o ka genome holoʻokoʻa, ʻike ʻia ʻo WGS, hiki ke hōʻike ʻia nā hoʻololi maʻamau a me nā ʻano like ʻole ma ka genome holoʻokoʻa me Single Nucleotide Polymorphism (SNP), Insertion Deletion (InDel), Structure variation (SV), a me Copy Number Variation (CNV). ).Hoʻokumu nā SV i kahi hapa nui o ka waihona hoʻololi ma mua o nā SNP a ʻoi aku ka nui o ka hopena i ka genome, he hopena koʻikoʻi i nā mea ola.ʻO ka heluhelu lōʻihi e hiki ai ke ʻike pololei i nā ʻāpana nui a me nā ʻano paʻakikī no ka mea ʻoi aku ka maʻalahi o ka heluhelu lōʻihi i ka hele ʻana o ka chromosomal ma luna o nā wahi paʻakikī e like me ka tandem repeats, GC/AT-rich regions, a me hyper-variable regions.
Papahana: Illumina, PacBio, Nanopore
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BMKMANU S1000 Spatial Transcriptome
Kū ka Spatial transcriptomics ma ke alo o ka ʻepekema ʻepekema, e hāʻawi ana i ka poʻe noiʻi e ʻimi i nā hiʻohiʻona hōʻike gene paʻakikī i loko o nā ʻiʻo me ka mālama ʻana i kā lākou pōʻaiapuni spatial.Ma waena o nā paepae like ʻole, ua hoʻomohala ʻo BMKGene i ka BMKManu S1000 Spatial Transcriptome Chip, e kaena ana i kahihoʻonui i ka hoʻonāo 5µM, a hiki i ka laulā subcellular, a hikinā hoʻonohonoho hoʻonā pae nui.ʻO ka puʻupuʻu S1000, e hōʻike ana ma kahi o 2 miliona mau kiko, hoʻohana i nā microwells i hoʻopaʻa ʻia me nā pahu i hoʻopiha ʻia me nā probes hopu hopu.Ua hoʻomākaukau ʻia kahi waihona cDNA, i hoʻonui ʻia me nā barcodes spatial, mai ka chip S1000 a ma hope i hoʻonohonoho ʻia ma ke kahua ʻo Illumina NovaSeq.ʻO ka hui pū ʻana o nā hōʻailona spatially barcoded a me nā UMI e hōʻoia i ka pololei a me ka kikoʻī o ka ʻikepili i hana ʻia.Aia ka hiʻohiʻona kūʻokoʻa o ka chip BMKManu S1000 i kona versatility, e hāʻawi ana i nā hoʻonohonoho hoʻonā nui-level i hiki ke hoʻopili maikaʻi ʻia i nā ʻāpana like ʻole a me nā pae kikoʻī.Hoʻonohonoho kēia adaptability i ka chip ma ke ʻano he koho maikaʻi loa no nā haʻawina transcriptomics spatial like ʻole, e hōʻoia ana i ka clustering spatial pololei me ka leo liʻiliʻi.
Ke hoʻohana nei i ka chip BMKManu S1000 a me nā ʻenehana transcriptomic spatial ʻē aʻe, hiki i nā mea noiʻi ke loaʻa ka ʻike maikaʻi aʻe o ka hoʻonohonoho spatial o nā cell a me nā pilina molekala paʻakikī e kū nei i loko o nā ʻiʻo, e hāʻawi ana i nā ʻike waiwai nui i nā mīkini i lalo o nā kaʻina hana ola i loko o nā ʻano ākea. oncology, neuroscience, developmental biology, immunology and botanical studies.
Papahana: BMKManu S1000 chip a me Illumina NovaSeq
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10x Genomics Visium Spatial Transcriptome
ʻO Spatial transcriptomics kahi ʻenehana ʻokiʻoki e hiki ai i nā mea noiʻi ke noiʻi i nā hiʻohiʻona hōʻike gene i loko o nā ʻiʻo i ka wā e mālama ana i kā lākou pōʻaiapili spatial.ʻO kahi kahua ikaika ma kēia kikowaena ʻo 10x Genomics Visium i hui pū ʻia me ke kaʻina Illumina.Aia ke kumumanaʻo o 10X Visium ma kahi puʻupuʻu kūikawā me kahi wahi hopu i koho ʻia kahi i waiho ʻia ai nā ʻāpana kiko.Aia kēia wahi hopu i nā wahi barcoded, e pili ana kēlā me kēia me kahi wahi kikoʻī i loko o ka ʻiʻo.Hoʻopaʻa inoa ʻia nā molekala RNA i hopu ʻia mai ka ʻiʻo me nā molecular identifiers (UMIs) i ka wā o ke kaʻina hana unuhi.Hāʻawi kēia mau wahi barcoded a me nā UMI i ka palapala ʻāina kikoʻī pololei a me ka helu ʻana o ka hōʻike gene ma kahi hoʻonā hoʻokahi.ʻO ka hui pū ʻana o nā hōʻailona spatially barcoded a me nā UMI e hōʻoia i ka pololei a me ka kikoʻī o ka ʻikepili i hana ʻia.Ma ka hoʻohana ʻana i kēia ʻenehana Spatial Transcriptomics, hiki i nā mea noiʻi ke loaʻa ka ʻike hohonu o ka hoʻonohonoho spatial o nā cell a me nā pilina molekala paʻakikī e kū nei i loko o nā ʻiʻo, e hāʻawi ana i nā ʻike koʻikoʻi i nā hana i lalo o nā kaʻina biological i nā ʻano he nui, me ka oncology, neuroscience, developmental biology, immunology. , a me nā haʻawina botanical.
Papahana: 10X Genomics Visium a me Illumina NovaSeq
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ʻO ka lōʻihi mRNA Sequencing-Nanopore
ʻOiai ʻo NGS-based mRNA sequencing e lawelawe ma ke ʻano he mea hana maʻamau no ka helu ʻana i ka huaʻōlelo gene, ʻo kona hilinaʻi ʻana i nā heluhelu pōkole e kaupalena i kona pono i nā loiloi transcriptomic paʻakikī.ʻO ka Nanopore sequencing, ma ka ʻaoʻao ʻē aʻe, hoʻohana i ka ʻenehana heluhelu lōʻihi, e hiki ai i ke kaʻina o nā transcripts mRNA holoʻokoʻa.Hoʻomaʻamaʻa kēia ala i ka ʻimi piha ʻana o ka splicing ʻē aʻe, nā fusions gene, poly-adenylation, a me ka helu ʻana o nā isoforms mRNA.
ʻO ke kaʻina hana Nanopore e hilinaʻi ana i nā hōʻailona uila i ka manawa maoli nanopore.Ke alakaʻi ʻia nei e nā polokina kaʻa, hoʻopaʻa ʻia ka DNA kaulalua i nā protein nanopore i hoʻokomo ʻia i loko o kahi biofilm, me ka wehe ʻana i kona hele ʻana ma ke kahawai nanopore ma lalo o kahi ʻokoʻa uila.ʻIke ʻia a hoʻokaʻawale ʻia nā hōʻailona uila i hana ʻia e nā kumu like ʻole ma ke kaula DNA i ka manawa maoli, e hoʻomaʻamaʻa i ka hoʻonohonoho nucleotide pololei a hoʻomau.ʻO kēia ala hou e lanakila ai i nā palena heluhelu pōkole a hāʻawi i kahi kahua hoʻoikaika no ka loiloi genomic paʻakikī, me nā haʻawina transcriptomic paʻakikī.
Papahana: Nanopore Promethion P48
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ʻO ke kaʻina mRNA lōʻihi piha -PacBio
ʻOiai ʻo NGS-based mRNA sequencing he mea hana maʻalahi no ka helu ʻana i ka hōʻike gene, ʻo kona hilinaʻi ʻana i nā heluhelu pōkole e kaupalena i kona hoʻohana ʻana i nā loiloi transcriptomic paʻakikī.ʻO PacBio sequencing (Iso-Seq), ma ka ʻaoʻao ʻē aʻe, hoʻohana i ka ʻenehana heluhelu lōʻihi, e hiki ai i ke kaʻina o nā transcripts mRNA lōʻihi.Hoʻomaʻamaʻa kēia ala i ka ʻimi piha ʻana o ka splicing ʻē aʻe, ka hui pū ʻana a me ka poly-adenylation ʻoiai ʻaʻole ia ke koho mua no ka helu ʻana i nā ʻano gene, ma muli o ka nui o ka ʻikepili i koi ʻia.
Ke hilinaʻi nei ka ʻenehana sequencing ʻo PacBio i ka hoʻopaʻa ʻana o ka molekala hoʻokahi, ka manawa maoli (SMRT), e hāʻawi ana i kahi pono kūʻokoʻa i ka hopu ʻana i nā transcript mRNA holoʻokoʻa.Hoʻohana ʻia kēia ala hou i ka hoʻohana ʻana i nā alakaʻi nalu mode zero (ZMWs), nā pūnāwai microfabricated e hiki ai ke nānā pono i ka hana DNA polymerase i ka wā o ka hoʻonohonoho ʻana.I loko o kēia mau ZMW, hoʻohui ka DNA polymerase o PacBio i kahi kaula hoʻohui o DNA, e hana ana i nā heluhelu lōʻihi e pili ana i ka holoʻokoʻa o nā transcripts mRNA.ʻO ka hana ʻo PacBio ma ke ʻano Circular Consensus sequencing (CCS) e hoʻonui i ka pololei ma o ka hoʻopili pinepine ʻana i ka mole like.Loaʻa ka pololei o nā heluhelu HiFi i hoʻohālikelike ʻia me NGS, e hāʻawi hou ana i kahi loiloi piha a hilinaʻi o nā hiʻohiʻona transcriptomic paʻakikī.Papahana: PacBio Sequel II
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Eukaryotic mRNA Sequencing-Illumina
Hāʻawi ka mRNA sequencing i ka hōʻike piha ʻana o nā transcripts mRNA āpau i loko o nā cell ma lalo o nā kūlana kikoʻī.Ke hana nei kēia ʻenehana ʻokiʻoki ma ke ʻano he mea hana ikaika, e wehe ana i nā profile hōʻike gene paʻakikī, nā hana gene, a me nā mīkini molekala e pili ana i nā kaʻina olaola like ʻole.Hoʻohana nui ʻia i ka noiʻi koʻikoʻi, nā diagnostics clinical, a me ka hoʻomohala ʻana i ka lāʻau lapaʻau, hāʻawi ʻo mRNA sequencing i nā ʻike i ka paʻakikī o ka dynamics cellular a me ka hoʻoponopono genetic.
Papahana: Illumina NovaSeq X
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Non-Reference based mRNA Sequencing-Illumina
Hāʻawi ka mRNA sequencing i ka hōʻike piha ʻana o nā transcripts mRNA āpau i loko o nā cell ma lalo o nā kūlana kikoʻī.Ke hana nei kēia ʻenehana ʻokiʻoki ma ke ʻano he mea hana ikaika, e wehe ana i nā profile hōʻike gene paʻakikī, nā hana gene, a me nā mīkini molekala e pili ana i nā kaʻina olaola like ʻole.Hoʻohana nui ʻia i ka noiʻi koʻikoʻi, nā diagnostics clinical, a me ka hoʻomohala ʻana i ka lāʻau lapaʻau, hāʻawi ʻo mRNA sequencing i nā ʻike i ka paʻakikī o ka dynamics cellular a me ka hoʻoponopono genetic.
Papahana: Illumina NovaSeq X
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Ka lōʻihi non-coding sequencing-Illumina
ʻO nā RNA lōʻihi ʻole-coding (lncRNAs) he mau RNA ka lōʻihi ma mua o 200 nucleotides i loaʻa ka liʻiliʻi liʻiliʻi coding a he mau mea koʻikoʻi i loko o ka RNA non-coding.Loaʻa i loko o ka nucleus a me ka cytoplasm, hoʻokani koʻikoʻi kēia mau RNA i ka epigenetic, transcriptional, a me ka hoʻoponopono post-transcriptional, e hōʻike ana i ko lākou koʻikoʻi i ka hana ʻana i nā kaʻina cellular a me ka molecular.ʻO ka LncRNA sequencing kahi mea hana ikaika i ka ʻokoʻa Cell, Ontogenesis, a me nā maʻi kanaka.
Papahana: Illumina NovaSeq
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Liʻiliʻi RNA sequencing-Illumina
ʻO nā molekole RNA liʻiliʻi (sRNA), ma lalo o 200 nucleotides ka lōʻihi, e komo pū me nā microRNAs (miRNAs), nā RNA interfering liʻiliʻi (siRNAs), a me nā RNA pili piwi (piRNAs).Ma waena o kēia mau mea, ʻo nā miRNA, ma kahi o 20-24 nucleotides ka lōʻihi, ʻike nui ʻia no kā lākou kuleana hoʻoponopono pivotal i nā kaʻina hana kelepona.Me nā hiʻohiʻona hōʻike kikoʻī kikoʻī a me ka pae, hōʻike nā miRNA i ka mālama kiʻekiʻe ma waena o nā ʻano like ʻole.
Papahana: Illumina NovaSeq
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circRNA sequencing-Illumina
ʻO ka hoʻopili ʻana i ka RNA circular (circRNA-seq) ʻo ia ka hoʻopili ʻana a me ka nānā ʻana i nā RNA pōʻai, kahi papa o nā molekala RNA i hana i nā puka puka pani ma muli o nā hanana splicing non-canonical, e hāʻawi ana i kēia mau RNA me ka hoʻonui paʻa.ʻOiai ua hōʻike ʻia kekahi mau circRNAs e hana e like me ka microRNA sponges, hoʻokaʻawale i nā microRNAs a pale iā lākou mai ka hoʻoponopono ʻana i kā lākou mau mRNA i manaʻo ʻia, hiki i nā circRNA ʻē aʻe ke launa pū me nā protein, hoʻololi i ka hōʻike gene, a i ʻole he kuleana i nā kaʻina kelepona.Hāʻawi ka hōʻike hōʻike circRNA i nā ʻike i nā kuleana hoʻoponopono o kēia mau molekala a me ko lākou koʻikoʻi i nā kaʻina kelepona like ʻole, nā pae hoʻomohala, a me nā kūlana maʻi, e hāʻawi ana i ka ʻike hohonu o ka paʻakikī o ka hoʻoponopono RNA ma ke ʻano o ka hōʻike gene.
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ʻO ke kaʻina transcriptome holoʻokoʻa - Illumina
Hāʻawi ka holo transcriptome holoʻokoʻa i kahi ala ākea i ka hoʻopili ʻana i nā molekole RNA like ʻole, e hoʻopili ana i nā coding (mRNA) a me nā RNA non-coding (lncRNA, circRNA, a me miRNA).Hoʻopili kēia ʻenehana i ka transcriptome piha o nā cell kikoʻī i ka manawa i hāʻawi ʻia, e ʻae ai i ka ʻike holoʻokoʻa o nā kaʻina kelepona.ʻIke ʻia hoʻi ʻo "total RNA sequencing," manaʻo ia e wehe i nā ʻoihana hoʻoponopono paʻakikī i ka pae transcriptome, e hiki ai i ka loiloi hohonu e like me ka hoʻokūkū RNA endogenous (ceRNA) a me ka loiloi RNA hui.Hōʻailona kēia i ka hana mua i ke ʻano hana, ʻoi aku ka wehe ʻana i nā ʻoihana hoʻoponopono e pili ana i nā pilina ceRNA circRNA-miRNA-mRNA.
