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BMKMANU S3000_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 S3000 Spatial Transcriptome Chip, e kaena ana i kahi hoʻonā i hoʻonui ʻia o 3.5µm, hiki i ka laulā subcellular, a hiki i nā hoʻonohonoho hoʻonā pae-nui. ʻO ka puʻupuʻu S3000, e hōʻike ana ma kahi o 4 miliona mau wahi, hoʻohana i nā microwells i hoʻopaʻa ʻia me nā pahu i hoʻopiha ʻia me nā probes hopu hopu spatially barcoded. Ua hoʻomākaukau ʻia kahi waihona cDNA, i hoʻonui ʻia me nā barcodes spatial, mai ka chip S3000 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. ʻO ka puʻupuʻu BMKManu S3000 he mea maʻalahi loa, e hāʻawi ana i nā hoʻonohonoho hoʻonā i nā pae he nui i hiki ke hoʻoponopono maikaʻi ʻia i nā ʻiʻo like ʻole a me nā pae kikoʻī i makemake ʻia. 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. ʻO ka hoʻohana ʻana i ka ʻenehana hoʻokaʻawale kelepona me BMKManu S3000 hiki ke hoʻokaʻawale i ka ʻikepili transcriptional i nā palena o nā cell, ka hopena i kahi loiloi i loaʻa ka manaʻo biological pololei. Eia kekahi, ʻoi aku ka maikaʻi o ka hoʻonā ʻana o S3000 i ka nui o nā genes a me nā UMI i ʻike ʻia i kēlā me kēia cell, e hiki ai i ka nānā pono ʻana i nā ʻano kikoʻī spatial a me ka hui ʻana o nā cell.
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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
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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
