-
ʻIkepili ʻAhahui Genome-wide
ʻO ka pahuhopu o Genome-Wide Association Studies (GWAS) ʻo ia ka ʻike ʻana i nā ʻano genetic (genotypes) i hoʻopili ʻia i nā ʻano kikoʻī (phenotypes). Ma ka nānā ʻana i nā hōʻailona genetic ma waena o ka genome holoʻokoʻa i kahi heluna nui o nā kānaka, hoʻonui ʻo GWAS i nā hui genotype-phenotype ma o ka helu helu helu kanaka. Loaʻa kēia ʻano hana i nā noi nui i ka noiʻi ʻana i nā maʻi kanaka a me ka ʻimi ʻana i nā genes hana e pili ana i nā ʻano paʻakikī o nā holoholona a mea kanu paha.
Ma BMKGENE, hāʻawi mākou i ʻelua ala no ke alakaʻi ʻana i ka GWAS ma nā heluna nui: hoʻohana i ka Whole-Genome Sequencing (WGS) a i ʻole ke koho ʻana i kahi ala hoʻohālikelike genome sequencing hōʻemi, ʻo ka Specific-Locus Amplified Fragment (SLAF) i kūkulu ʻia i loko o ka hale. ʻOiai ua kūpono ka WGS i nā genome liʻiliʻi, ua puka mai ʻo SLAF ma ke ʻano he kumu kūʻai kūpono no ke aʻo ʻana i nā heluna nui me nā genomes lōʻihi, e hōʻemi pono ana i nā kumukūʻai sequencing, ʻoiai e hōʻoiaʻiʻo ana i kahi kiʻekiʻe o ka ʻike ʻana i ka hōʻailona genetic.
-
Hoʻokahi-nuclei RNA Sequencing
ʻO ka hoʻomohala ʻana o ka hoʻopaʻa ʻana i ke kelepona hoʻokahi a me nā ʻenehana kūkulu hale waihona puke, i hui pū ʻia me ke kaʻina hana kiʻekiʻe, ua hoʻololi i nā haʻawina hōʻike gene ma ka pae kelepona. Hāʻawi kēia holomua i ka loiloi hohonu a ʻoi aku ka hohonu o nā heluna cell paʻakikī, e lanakila ana i nā palena e pili ana i ka hōʻike ʻana o nā gene ma luna o nā cell āpau a mālama i ka heterogeneity maoli i loko o kēia mau heluna. ʻOiai ʻo ka hoʻokaʻina ʻana o ka RNA cell hoʻokahi (scRNA-seq) loaʻa nā pōmaikaʻi hiki ʻole ke hōʻole ʻia, loaʻa iā ia nā paʻakikī i kekahi mau kiko kahi i paʻakikī ai ka hana ʻana o ka hoʻokuʻu ʻana i hoʻokahi cell a koi i nā laʻana hou. Ma BMKGene, hoʻoponopono mākou i kēia pilikia ma ka hāʻawi ʻana i ka hoʻokaʻina RNA nucleus hoʻokahi (snRNA-seq) me ka hoʻohana ʻana i ka ʻenehana 10X Genomics Chromium hou. Hoʻonui kēia ala i ka spectrum o nā laʻana i hiki ke hoʻopaʻa ʻia i ka transcriptome analysis ma ka pae hoʻokahi.
Hoʻokō ʻia ka hoʻokaʻawale ʻana o ka nuclei ma o ka puʻupuʻu 10X Genomics Chromium hou, e hōʻike ana i kahi ʻōnaehana microfluidics ʻewalu-kanela me nā ala ʻelua. I loko o kēia ʻōnaehana, ua hoʻopili ʻia nā pahu gel e hoʻokomo ana i nā barcodes, primers, enzymes, a me kahi nucleus hoʻokahi i loko o nā kulu aila nui nanoliter, e hana ana i ka Gel Bead-in-Emulsion (GEM). Ma hope o ka hoʻokumu ʻana o GEM, hoʻokuʻu ʻia ka cell lysis a me ka barcode i loko o kēlā me kēia GEM. Ma hope mai, hoʻololi ʻia nā molekala mRNA i loko o nā cDNA, e hoʻopili ana i nā barcode 10X a me nā Unique Molecular Identifiers (UMIs). Hoʻokomo ʻia kēia mau cDNA i ke kūkulu ʻana i ka waihona sequencing maʻamau, e hoʻomaʻamaʻa i ka ʻimi ikaika a paʻa i nā profile hōʻike gene ma ka pae hoʻokahi.
Papahana: 10× Genomics Chromium a me Illumina NovaSeq Platform
-
Mea kanu/holoholona holoʻokoʻa Genome Sequencing
ʻO ka Whole Genome Sequencing (WGS), i ʻike ʻia ʻo resequencing, pili i ka hoʻonohonoho genome holoʻokoʻa o nā kānaka like ʻole o nā ʻano me nā genome kuhikuhi. Ma kēia kumu, hiki ke ʻike hou ʻia nā ʻokoʻa genomic o nā kānaka a i ʻole nā kānaka. Hiki i ka WGS ke hoʻomaopopo i ka Single Nucleotide Polymorphism (SNP), Insertion Deletion (InDel), Structure variation (SV), a me Copy Number Variation (CNV). Aia nā SV i kahi hapa nui o ka waihona hoʻololi ma mua o nā SNP a ʻoi aku ka hopena o ka genome, e pili nui ana i nā mea ola. ʻOiai ʻoi aku ka maikaʻi o ka resequencing heluhelu pōkole i ka ʻike ʻana i nā SNP a me InDels, hiki i ka resequencing heluhelu lōʻihi ke ʻike pololei i nā ʻāpana nui a me nā ʻano paʻakikī.
-
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
-
ʻO ka lōʻihi mRNA Sequencing-Nanopore
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 pono i nā loiloi transcriptomic paʻakikī. Ma ka ʻaoʻao ʻē aʻe, hoʻohana ʻo nanopore sequencing i ka ʻenehana heluhelu lōʻihi, e hiki ai i ke kaʻina o nā transcripts mRNA lōʻihi piha. 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 Nanopore sequencing, kahi ala e hilinaʻi ai i nā nanopore hoʻokahi-molele i ka manawa maoli, hāʻawi i nā hopena i ka manawa maoli. 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 ʻokoʻa 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, me nā hopena koke.
Papahana: Nanopore PromethION 48
-
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. Ma ka ʻaoʻao ʻē aʻe, hoʻohana ʻo PacBio sequencing (Iso-Seq) 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 ʻokoʻa, ka hui pū ʻana, a me ka poly-adenylation. Eia nō naʻe, aia kekahi mau koho ʻē aʻe no ka helu ʻana i nā huaʻōlelo 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) a me nā pūnāwai microfabricated e hiki ai ke nānā i ka manawa maoli o ka hana DNA polymerase i ka wā o ka sequencing. 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. Hoʻonui ka hana ʻo PacBio ma ke ʻano Circular Consensus sequencing (CCS) i ka pololei ma o ka hoʻopili pinepine ʻana i ka molela 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; PacBio Revio
-
Eukaryotic mRNA Sequencing-NGS
ʻO ka mRNA sequencing, kahi ʻenehana loea, hāʻawi i ka mana i ka hoʻopili piha ʻana o nā transcripts mRNA āpau i loko o nā cell ma lalo o nā kūlana kikoʻī. Me kāna mau noi ākea, ua wehe kēia mea hana ʻokiʻoki i nā profile hōʻike gene paʻakikī, nā ʻano hana gene, a me nā mīkini molekala e pili ana i nā kaʻina hana olaola like ʻole. Hoʻohana nui ʻia i ka noiʻi kumu, 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, e hoʻāla ana i ka ʻike e pili ana i kona hiki i nā ʻano like ʻole.
Papahana: Illumina NovaSeq X; DNSSEQ-T7
-
Non-Reference based mRNA Sequencing-NGS
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 moʻolelo hōʻikeʻike gene paʻakikī, nā ʻano hana gene, a me nā mīkini molekala e pili ana i nā kaʻina hana 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 e pili ana i ka intricacies o ka cellular dynamics a me ka hoʻoponopono genetic.
Papahana: Illumina NovaSeq X; DNSSEQ-T7
-
Long Non-coding Sequencing-Illumina
ʻOi aku ka lōʻihi o nā RNA non-coding (lncRNAs) 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
-
Liʻiliʻi RNA Sequencing-Illumina
ʻO nā molekole RNA liʻiliʻi (sRNA), 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 18-25 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
-
CircRNA Sequencing-Illumina
ʻO ka circular RNA sequencing (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 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 i ka pōʻaiapili o ka hōʻike gene.
-
ʻO ke Kaʻina Transcriptome holoʻokoʻa - Illumina
Hāʻawi ka holo transcriptome holoʻokoʻa i kahi ala holoʻokoʻa i ka hoʻopili ʻana i nā molekole RNA like ʻole, e hoʻopili ana i ka coding (mRNA) a me nā RNA non-coding (lncRNA, circRNA, a me miRNA). Hoʻopili kēia ʻenehana i ka transcriptome holoʻokoʻa o nā cell kikoʻī i ka manawa i hāʻawi ʻia, e ʻae ai i ka ʻike holoʻokoʻa o nā kaʻina kelepona. ʻIke ʻia ʻo "total RNA sequencing," manaʻo ia e wehe i nā ʻoihana hoʻoponopono koʻikoʻi ma ka pae transcriptome, e hiki ai i ka loiloi hohonu e like me ka 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.
-
ʻO Chromatin Immunoprecipitation Sequencing (ChIP-seq)
ʻO ka Chromatin Immunoprecipitation (CHIP) kahi hana e hoʻohana ai i nā antibodies e hoʻonui i nā protein paʻa DNA a me kā lākou mau pahuhopu genomics. ʻO kona hoʻohui pū ʻana me NGS e hiki ai i ka genome-wide profiling o nā pahuhopu DNA e pili ana me ka hoʻololi histone, nā kumu transcription, a me nā protein paʻa DNA ʻē aʻe. Hiki i kēia ala hoʻoikaika ke hoʻohālikelike i nā pūnaewele paʻa ma waena o nā ʻano cell like ʻole, nā kiko, a i ʻole nā kūlana. Loaʻa nā noi a ChIP-Seq mai ke aʻo ʻana i ka hoʻoponopono transcriptional a me nā ala hoʻomohala i ka wehewehe ʻana i nā mīkini maʻi, e lilo ia i mea hana pono no ka hoʻomaopopo ʻana i nā ʻāina hoʻoponopono genomic a me ka holomua ʻana i nā ʻike therapeutic.
Papahana: Illumina NovaSeq
