Lihlahisoa

Sequloana se le seng sa RNA Sequencing

Single-nucleus RNA Sequencing Featured Image

Tsoelo-pele ea ho nka sele e le 'ngoe le mokhoa oa ho haha laebrari ka bomong ho kopanngoa le tatellano e phahameng e lumellang lithuto tsa liphatsa tsa lefutso motheong oa sele-by-cell.E etsa hore ho be le tlhahlobo e tebileng le e felletseng ea litsamaiso tsa lisele tse rarahaneng, moo haholo-holo e qobang ho pata liphapang tsa bona ka ho nka karolelano ea lisele tsohle.

Leha ho le joalo, lisele tse ling ha li tšoanelehe ho etsoa ho emisoa ha sele e le 'ngoe, kahoo mekhoa e meng ea ho lokisa sampole - ho ntšoa ha nucleus ho tsoa liseleng hoa hlokahala, ke hore, nucleus e ntšoa ka kotloloho ho lisele kapa lisele ebe e lokisetsoa ho emisoa ha nucleus e le 'ngoe bakeng sa motho a le mong. tatellano ea lisele.

BMK e fana ka ts'ebeletso ea tatellano ea 10× Genomics ChromiumTM e thehiloeng ho sele e le 'ngoe ea RNA.Ts'ebeletso ena e sebelisitsoe haholo lithutong tse amanang le mafu, joalo ka phapang ea lisele tsa 'mele oa ho itšireletsa mafung, tumor heterogeneity, nts'etsopele ea lisele, joalo-joalo.

Spatial transcriptome chip: 10 × Genomics

Sethala: Illumina NovaSeq Platform

Iteanye le rona

Lintlha tsa Tšebeletso

Bioinformatics

Demo Results

Molao-motheo oa theknoloji

Ho aroloa ha li-nuclei ho finyelloa ke 10 × Genomics Chromium TM, e nang le tsamaiso ea microfluidics e nang le mechine e robeli e nang le ho tšela habeli.Tsamaisong ena, lifaha tsa gel tse nang le li-barcode le primer, li-enzyme le nucleus e le 'ngoe li kenngoa ka lerotholi la oli ea nanoliter, e hlahisang Gel Bead-in-Emulsion (GEM).Hang ha GEM e thehoa, lisele tsa lysis le ho lokolloa ha barcode li etsoa ho GEM ka 'ngoe.mRNA e fetisetsoa ka morao ho limolek'hule tsa cDNA tse nang le 10 × barcodes le UMI, tse tlas'a taolo ea kaho e tloaelehileng ea tatellano ea laeborari.

Lithane ha li tšoanelehe bakeng sa tokiso ea ho emisoa ha sele e le 'ngoe

Sele/ Sehlahisoa	Lebaka
Lithipa tse hatselitsoeng tse sa nchafalang	Ha e khone ho fumana mekhatlo e mecha kapa e bolokiloeng nako e telele
Lisele tsa mesifa, Megakaryocyte, Mafura ...	Bophahamo ba sele bo boholo haholo ho kena seletsa
Sebete...	E robeha haholo hore e ka robeha, ha e khone ho khetholla lisele tse le 'ngoe
Neuron cell, boko…	Ho utloisisa haholoanyane, ho bonolo ho hatella, ho tla fetola liphetho tsa tatellano
Makhopho, Thyroid ...	E ruile ka li-enzyme tsa endogenous, tse amang tlhahiso ea ho emisoa ha sele e le 'ngoe

Nucleus e le 'ngoe vs Lesele e le 'ngoe

Nucleus e le 'ngoe	Sele e le 'ngoe
Sekhahla se se nang moeli sa sele	Bophara ba sele: 10-40 μm
Thepa e ka ba lesela le leqhoa	Boitsebiso bo tlameha ho ba lisele tse ncha
Khatello e tlase ea lisele tse leqhoa	Kalafo ea enzyme e ka baka khatello ea maikutlo ea lisele
Ha ho na lisele tse khubelu tsa mali tse lokelang ho tlosoa	Lisele tse khubelu tsa mali li hloka ho tlosoa
Nuclear e hlahisa bioinformation	Sele eohle e hlahisa litaba tsa bophelo

Litlhaloso tsa Tšebeletso

Laebrari

Leano la tatellano

Bophahamo ba data

Litlhoko tsa Mohlala

Tishu

10 × Laeborari ea li-nuclei tse le 'ngoe tsa Genomics

10x Genomics -Illumina PE150

100,000 e baloa/lisele hoo e ka bang.100-200 Gb

Nomoro ea sele: >2×10⁵

Khokahano ea sele.ho 700-1,200 sele / μL

≥ 200 mg

Bakeng sa lintlha tse ling mabapi le tataiso ea boitokisetso ea mohlala le ts'ebetso ea ts'ebeletso, ka kopo ikutloe u lokolohile ho bua le aBMKGENE setsebi