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条形 fu'a-03

Oloa

Fa'asologa o le RNA nucleus e tasi

O le aga'i i luma i le pu'eina o sela e tasi ma faiga fau faletusi ta'ito'atasi e tu'ufa'atasia ma le fa'asologa maualuga e mafai ai ona su'esu'eina fa'amatalaga o kenera i luga ole sela ma le sela.E mafai ai se su'esu'ega loloto ma atoatoa i luga o le faitau aofa'i o sela lavelave, lea e 'alo'ese ai le fa'amamaluina o lo latou fete'ena'i e ala i le ave'ese o sela uma.

Ae ui i lea, o nisi sela e le talafeagai ona faia e avea ma suspension e tasi-cell, o lea o isi metotia faʻataʻitaʻiga sauniuni - e manaʻomia le faʻaogaina o le nucleus mai tissues, o lona uiga, o le nucleus e sau saʻo mai le masini poʻo le cell ma saunia i totonu o le tasi-nucleus suspension mo le tasi- fa'asologa o sela.

BMK e tuʻuina atu le 10 × Genomics Chromium TM faʻavae tasi-cell RNA sequencing auaunaga.O lenei 'au'aunaga ua fa'aaogaina lautele i su'esu'ega i su'esu'ega fa'ama'i, e pei o le fa'avasegaina o le tino puipuia, tumo heterogeneity, fa'atupuina o le tino, ma isi.

Vaila'au fa'asologa fa'apitoa: 10× Genomics

Tulaga: Illumina NovaSeq Platform


Au'aunaga Fa'amatalaga

Bioinformatics

I'uga Fa'aaliga

mataupu faavae fa'apitoa

O le vavaeeseina o le nuclei e ausia e le 10 × Genomics Chromium TM, lea e aofia ai le microfluidics system e valu-channel faʻatasi ai ma faʻasalalauga faʻalua.I lenei faiga, o se pulu gel ma barcodes ma primer, enzymes ma se tasi nucleus o loo encapsulated i nanoliter-sized suauu mataua, e gaosia Gel Bead-i-Emulsion (GEM).O le taimi lava e fausia ai le GEM, o le cell lysis ma le tatalaina o barcodes e faia i GEM taitasi.O le mRNA o loʻo faʻaliliuina i totonu o mole mole cDNA ma 10x barcodes ma UMI, lea e faʻatatau i le faʻasologa masani o le fausiaina o faletusi.

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E le talafeagai le i'u mo le tapenaga o le fa'amalologa e tasi

Cell / Tissue

Mafuaaga

Fa'a'aisa le mea fou

Le mafai ona maua ni fa'alapotopotoga fou pe ua leva ona fa'asaoina

Sel maso, Megakaryocyte, Ga'o…

O le lautele o le sela e telē tele e ulufale ai i le meafaigaluega

ate…

E matua maaleale e malepe, e le mafai ona iloa se tasi o sela

Neuron cell, Fai'ai…

E sili atu le maaleale, faigofie ona faʻamalosia, o le a suia ai le faʻasologa o taunuuga

Pancreas, thyroid…

Mauoa i enzymes endogenous, e a'afia ai le gaosiga o le fa'amalologa o le sela e tasi

Nucleus tasi vs Selula tasi

Nucleus tasi

sela tasi

E leai se lautele ole sela

Ole lapoa ole sela: 10-40 μm

O mea e mafai ona fa'aisa aisa

O mea e tatau ona fou

Maualalo le fa'amamafa o sela fa'a'aisa

O le togafitiga ole enzyme e mafai ona mafua ai le fa'afitauli o le tino

E leai se toto mumu e tatau ona aveese

E tatau ona aveese sela mumu toto

Nuclear fa'aalia bioinformation

O le sela atoa e faʻaalia bioinformation

Fa'amatalaga Auaunaga

Faletusi

Fuafuaga fa'asologa

Volume Fa'amatalaga

Fa'ata'ita'iga Manaoga

Tisu

10× Genomics tasi-nuclei faletusi

10x Genomics -Illumina PE150

100,000 faitau/selau pe tusa.100-200 Gb

Numera sela: >2× 105

Cell conc.ile 700-1,200 cell/μL

≥ 200 mg

Mo nisi fa'amatalaga i fa'ata'ita'iga tapenaga ta'iala ma au'aunaga, fa'amolemole lagona le saoloto e talanoa i a

Auala o Galuega Au'aunaga

Fa'ata'ita'iga QC

Fa'ata'ita'iga mamanu

fa'ata'ita'iga tu'uina atu

Fa'ata'ita'iga tu'uina atu

faaniukilia.faamalolo

Nuclei vavaeeseina

Sauniuniga Faletusi

Fauga faletusi

Fa'asologa

Fa'asologa

Iloiloga fa'amaumauga

Iloiloga fa'amaumauga

A uma le fa'atauga Auaunaga

Au'aunaga pe a uma ona fa'atau


  • Muamua:
  • Sosoo ai:

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    1.Spot fa'aputuina

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    2. Fa'ailoga fa'ailoga abundance fa'apipi'i fa'afanua vevela

    wps_doc_113. Fausia le fa'asoaina o kenera i fuifui eseese

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    4.Cell trajectory analysis/pseudotime

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