Single- nuxi rna seequing
Hana pāhana
Loaʻa ka makemake o nā nuku o nuclei e 10 × genomiss chromium ™, e like me nā'āpana heʻumikūmālua o nā papaʻelua. Ma kēia'ōnaehana, he Gel Boads me nā barcodes a me nā mea nui, e hoʻonuiʻia nā envymes a me kahi o ka ailaʻaila gel bided. Ua hanaʻiaʻo Gem, i hanaʻiaʻo Will Lysis a hoʻokuʻu i nā barcodes i hana i kēlā me kēia gem. Ua hoʻohuliʻiaʻo Mrna i nā mole cdna molecules me 10
Nā hiʻohiʻona
● E hoʻomākaukau ana i ka ukuʻana o hoʻokahi-nucle
● hoʻokumu i ka gel bel beed-in-emulsion (gem) i ukaliʻia e cdna synthesis
● Ua hoʻoukaʻia kēlā me kēia bead i kahi gem me nā primers i hoʻohuiʻia i 4 mau'āpana:
Poly (dt) huelo no mrna priming a me cynthesis,
ʻO kaʻike molecular Unique (UMI) e hoʻoponopono i nā bias amplification
10X bearde
Ke hoʻopaʻa nei i keʻano o ka helu helu helu 1 stequence primer
Loaʻa
ʻO keʻano hoʻokahi-nucleus RNA e hele ana i nā palena o nā palena o ka cenent-cenent-cenenting, e hiki ai:
●ʻO ka hoʻohanaʻana o nā kiʻi o nā brozen aʻaʻole wale wale nō i kaupalenaʻia i nā hiʻohiʻona hou
● He haʻahaʻa haʻahaʻa o nā chells brozen i ka wā e hoʻohālikelikeʻia ai ka hanaʻana i nāʻikepili hou i keʻano o nā mea kākau
●ʻAʻole pono no ka laweʻana i ka neʻeʻana o nā'āpana kokoʻulaʻula
● Nā Kūlana Kūʻai Kūʻai
● Acray nui o nā hiʻohiʻona e pono ai no ka nānāʻana, me ka paʻakikī a me ka Fragile Tissue Type a iʻole ka lukuʻana i ka wā o ka cissue
Nā'āpanaʻaʻole hiki ke loiloiʻia e nā cequing hoʻokahi-cequing a loaʻa iā ia no ka geque rna secequing.
| Cell / tissue | O ke kumu |
| ʻO nā kiko o Unfresh Frozen | ʻAʻole hiki ke loaʻa i nā hui hou a lōʻihi paha |
| Nā Cell Celcle, Megakyytete, momona ... | He nui loa ka helu cell diameter e komo i ka mea hana |
| Lio ... | ʻO Fragile e wāwahi,ʻaʻole hiki ke hoʻokaʻawale i nā'āpana hoʻokahi |
| Neuon Cell, Fain ... | ʻOi aku ka paʻakikī, maʻalahi ke kaumaha, e hoʻololi i nā hopena seequing |
| PACHORS, Kaukana ... | Waiwai ma nā enzyomous endyomus, e pili ana i ka hanaʻana o keʻano o ke kaomiʻana i ke kelepona hoʻokahi |
Hoʻokahi-nucleus vs cell-cell
| ʻO nāʻano likeʻole | Hoʻokahi-cell |
| ʻO keʻano o ke kelepona kūʻaiʻole | ʻO ka cell ka cell: 10-40 μm |
| Hiki i ka mea ke lilo i kaʻili | Pono nā kumuwaiwai hou |
| Haʻahaʻa haʻahaʻa o nā cell curzen | ʻO Enzyme ka hoʻomaʻamaʻa e hōʻeha i keʻano o keʻano |
| ʻAʻole pono e laweʻia nā wahi kokoʻulaʻula | Pono e laweʻia nā pahu kokoʻulaʻula |
| Hōʻikeʻo nukala i ka bioinformation | Hōʻike ka cell holoʻokoʻa i ka bioinformation |
Nā kiko'ī
| Nā koi koi | Puka Hanau | Serenencing hoʻolālā | Ua'ōleloʻia nāʻikepili | Honua mālamalama |
| Holoholona hihiu ≥ 200 mg Mea kanu lāʻau ≥ hola 100 mg | 10x genoms sl cdna ka hale waihona puke | Hoʻomālamalama pe150 | Ua heluheluʻo 100k Pe i kēlā me kēia cell (100-200 GB) | ʻO 700-1200 nuclei / μl a me nuclei integrity i nānāʻia ma lalo o ka microscope |
No nā kiko'ī hou aku ma nā kiko'ī o ka uku hoʻomākaukau a me ka lawelaweʻana, eʻoluʻolu e kamaʻilio e kamaʻilio meMele Kalikimaka
E holo ka lawelawe lawelawe
Komo i ka loiloi ma lalo nei:
● Hoʻonaʻauao maikaʻi:ʻO ka helu o nā cell, kaʻike a me kaʻike pololeiʻana o nā cell, rna molecules
● Inner Sport Sportis:
Cell clustering a me ka hoʻopiliʻana
ʻO nā hōʻike hōʻike hōʻikeʻokoʻa
Hana hana a me ka hoʻonuiʻana i nā degs cluster
● Kekuhi-Kūlana:
Hui me nā ikepili
ʻO nā hōʻike hōʻike hōʻikeʻokoʻa: kaʻikeʻana i nā degs i nā hui
Hana hana a me ka hoʻonuiʻana o nā papa hana o nā hui
● loiloi loiloi:
Ka loiloi cycle cycle
ʻO ka loiloi Pseudotime
ʻO ka loiloi kamaʻilio kelepona (kelepona)
Ua hoʻokumuʻo Genest i ka loiloi enrichment (GSEA)
Inner-sample ka loiloi
CELLSING CLUSTERING:
ʻO ka hōʻike hōʻikeʻokoʻa: nā degs degs
Ka loiloi hui waena
ʻO nā hōʻike hōʻikeʻokoʻa
Hoʻolaha holomua:
PSEDOTIME i hōʻikeʻia:
E nānā i nā kelepona
Eʻimi i nā holomua i hoʻomaikaʻiʻia e BMKGEN SIGNUSE-nuclenus RNA stequence Services e 10x chromium i kēia mau hōʻike hōʻikeʻia:
Wang, L. et al. (2021) 'o ke kelepona a me ke kelepona hana kākau inoa inoa i hōʻikeʻia i ka pae'āina o ka paila o ka ling i ka stegoma retacterbation asthma.Nā hana o ka lāhui aupuni o nāʻepekema oʻAmelika Hui PūʻIa oʻAmelika Hui PūʻIa, 118 (2), p. E200590118. Doi: 10.1073 / Pnas.200590118
Zheng, H. et al. (2022)ʻO nā mea noho ma ka immunology, 13, p. 879824. Doi: 10.3389 / Fimmu.20222.879824 / Bibtex.
Tian, H. et al. (2023) 'ONT-cell-cell-cell-cell cellcritenastome amerogenity a me nā pane hou o nā lepatotes ma hope o ka floundiet (paratichthys Olivacties)'.Paulol, 566, p. 739238. Doi: 10.1016 / J.Angacature.2023.739238.
Yua, y. et al. (2023) '' Acdynamic Therapy e hoʻomaikaʻi i ka hopena o ka hopena o nā ISMUNE VISICTOR i nā mea maʻi ATI-TUSTIC.ʻO Gasric Cancer, 26 (5), PP. 798-813. Doi: 10.1007 / S10120-023-01409-x / Metrics.
