Single- nuclei rna sequencing
Tekinoroji Scheme
Kuzviparadzanisa kweNuclei kunowanikwa ne 10 × genomics Chromefics ™, iyo inosanganisira sere-chiteshi microfluidics system ine maviri kuyambuka. Mune ino system, iyo gel beads ine barcode uye primer, enzymes uye imwe nucleus yakadzvanywa muNaneliter-saizi yekudonha kwemafuta, kugadzira gel bead-in-emulsion (gem). Kamwe gem yakaumbwa, cell lyiss uye kusunungurwa kweBarcode anoitwa mune yega yega gem. MrNN yakatsaurwa yakanyorwa muCDN MOLECULES ne10
Zvimiro
● Kugadzirira kusarudzika-nuclei kumiswa kubva kune chando tishu
● Kuumbwa kweGel Bead-in-emulsion (gem) ichiteverwa neCDNA SYNTHESIS
● Bheji rega rega mune gem inotakura zvimedu zvakanyorwa zvikamu zvina zvikamu:
poly (dt) muswe kune mrna prnaing uye cdna synthesis,
Yakasarudzika molecular identifier (UMI) kuti aruramise maAmplization bias
10x barcode
Kusunga kutevedzana kwechidimbu kuverenga 1 sequencing Primer
Zvakanakira
Imwe-nucusus rna inotsevha inokwanisika zvisingakwanisi kusarudzika kweimwe-cell rna kutsakatika, kugonesa:
● Iko kushandiswa kwemasampuji echando uye kwete kungogumira kune matsva samples
.
● Hapana chikonzero chekubviswa kwekutanga kwemasero matsvuku eropa
● Unelimited cell Diameter
● Kurongeka kwakakura kwemasampuli anokodzera kuongororwa, kusanganisira mitezo yakaoma uye isina kusimba yemhando yekunyepedzera kune cell clumping kana kuparadzwa panguva yekuparadzaniswa kwetissue
Samples isingagoni kuongororwa neyakaenzana-cell rna sequencing uye yakakodzera kune imwe chete nuclei rna yekuseka:
| Cell / tishu | Chikonzero |
| Unfresh chando tishu | Kutadza kuwana matsva kana marefu-akachengetedzwa masangano |
| Mushambo Cell, Megakararyocyte, mafuta ... | Cell diameter yakakura kupinda muchiridzwa |
| Chiropa ... | Kunyanya kusasimba kuputsa, kutadza kusiyanisa maseru asina kuroora |
| Neuron Cell, Brain ... | Zvakanyanya kungwarira, nyore kushushikana, kuchachinja goquencing mhinduro |
| Pancreas, Thyroid ... | Kupfuma mumagumo enzymes, inokanganisa kugadzirwa kwekumiswa kwechechi imwe chete |
Single-nuclus vs sero-sero
| Single-nucleus | Sero-sero |
| Isina muganho cell diameter | Cell Diameter: 10-40 μm |
| Izvo zvinyorwa zvinogona kuve zvando tishu | Izvo zvinyorwa zvinofanirwa kunge zviri matsva tishu |
| Kushushikana kwakaderera kwemasero echando | Enzyme kurapwa kunogona kukonzera kusagadzikana kwesero kuita |
| Hapana masero matsvuku eropa anofanirwa kubviswa | Matsvuku eropa anoda kubviswa |
| Nyukireya inoratidza bioinformation | Sero rese rinoratidza bioinformation |
Zvinyorwa
| Zvinodiwa Muenzaniso | Raibhurari | Kuongorora zano | Data yakakurudzirwa | Mhando yekudzora |
| Mhuka Tishu】 200 mg Chirimwa tishu ± 400 mg | 10x genomics sn cdna library | Illumina Pe150 | 100k Pe inoverengwa pasero (100-200 GB) | 700-1200 Nuclei / μl uye nuclei kutendeka kwaanoona pasi peMicroscope |
Kuti uwane mamwe mashoko pane sampu yekugadzirira kutungamira uye sevhisi yebasa, ndapota inzwa wakasununguka kutaura neBMKGENE NCEELT
Basa rebasa rinoyerera
Inosanganisira kuongororwa kunotevera:
● Kugadziriswa Kwemhando: Nhamba yemasero, gene kuona, kuzivikanwa kwakaringana kwemasero, rna molecules uye kujekesa kwehuwandu
● Remukati Sample Anquiss:
Cell clustering uye cluster annotation
Kusiyanisa kutaura kwezasi: Kuzivikanwa kweDGS mumasumbu
Inoshanda Information uye Kufumisa Kwemasumbu Degs
● Inter-boka kuongorora:
Kusanganisa kwedata
Kusiyanisa Expection Express kuongororwa: Kuzivikanwa kweDGGs mumapoka
Inoshanda Ingationation uye Kufumisa Kwemapoka Degs
● Kuongororwa kwedekmiso:
Cle cycle kuongorora
Pseudotime Ongororo
Cell kutaurirana kuongororwa (cellphonedb)
Gene Yakagadzika Kuongorora Kwekucherekedza (Gsea)
Yemukati-samplu yekuongorora
Cell inosimbisa:
Kusiyanisa Express Express Kuongorora: Cluster Degs
Inter-Boka Ongororo
Kusiyanisa Express Express Kuongorora: Boka Dondo
Kuongorora Kwekutanga:
Pseudotime kuongororwa:
Cle Cycle Ongororo:
Ongorora kufambira mberi kwakabatanidzwa neBMKGE 'NULLULUS RNA SEQUEUS SEASS NE10X Chromium mune idzi dzakaratidza mabhuku:
Wang, l. et al. . (Mafambiro eNational Academy weScience yeUnited States of America, 118 (2), p. E2005590118. DOI: 10.1073 / Pnas.2005590118
Zheng, H. Et Al. . "Mberiers muIntology, 13, p. 879824. Doi: 10.3389 / fimmu.2022.879824 / Bibtex.
Tian, H. Et Al. .Aquaculture, 566, p. 739238. DII: 10.1016 / J.Aquaculture.2023.738238.
Yu, Y. et al. .Gastric Cancer, 26 (5), mapeji 798-813. Doi: 10.1007 / S10120-023-01409-x / metrics.
