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Zogulitsa

Single-nucleus RNA Sequencing

Kutsogola kwa kujambula kwa selo limodzi ndi njira yopangira laibulale yamunthu payekha kuphatikiza ndi kutsata kopitilira muyeso kumathandizira maphunziro amtundu wa gene pa cell-by-cell.Imathandizira kusanthula kozama komanso kokwanira pama cell ovuta, momwe imapewa kubisala kusiyanasiyana kwawo potenga pafupifupi ma cell onse.

Komabe, ma cell ena sali oyenera kupangidwa kuti akhale kuyimitsidwa kwa selo limodzi, kotero njira zina zokonzekera zitsanzo - kuchotsa nyukiliya kuchokera ku minofu kumafunika, ndiko kuti, nyukiliya imachotsedwa mwachindunji ku minofu kapena selo ndikukonzedwa kuti ikhale imodzi-nucleus kuyimitsidwa kwa single- kutsatizana kwa ma cell.

BMK imapereka 10 × Genomics Chromium TM yochokera ku cell single RNA sequencing service.Utumikiwu wagwiritsidwa ntchito kwambiri pamaphunziro okhudzana ndi matenda, monga kusiyanitsa kwa maselo a chitetezo chamthupi, chotupa chosiyana, kukula kwa minofu, ndi zina zambiri.

Spatial transcriptome chip: 10 × Genomics

Pulatifomu: Illumina NovaSeq Platform


Tsatanetsatane wa Utumiki

Bioinformatics

Zotsatira za Demo

mfundo zaukadaulo

Kudzipatula kwa nuclei kumatheka ndi 10 × Genomics Chromium TM, yomwe ili ndi makina asanu ndi atatu a microfluidics omwe ali ndi maulendo awiri.M'dongosolo lino, mikanda ya gel yokhala ndi barcode ndi primer, ma enzymes ndi phata limodzi zimayikidwa mu dontho lamafuta la nanoliter, ndikupanga Gel Bead-in-Emulsion (GEM).GEM ikapangidwa, cell lysis ndi kutulutsidwa kwa barcode kumachitika mu GEM iliyonse.mRNA amalembedwanso m'mamolekyu a cDNA okhala ndi ma barcode 10 × ndi UMI, omwe amayang'aniridwanso ndi kachitidwe kotsatizana ka library.

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Minofu si oyenera single selo kuyimitsidwa kukonzekera

Selo / Thupi

Chifukwa

Minofu yowumitsidwa yatsopano

Simungathe kupeza mabungwe atsopano kapena osungidwa kwanthawi yayitali

Maselo a minofu, Megakaryocyte, Mafuta…

Ma cell awiri ndi akulu kwambiri kuti alowe mu chida

Chiwindi…

Ndizosalimba kwambiri kuti zitha kusweka, sizimatha kusiyanitsa maselo amodzi

Maselo a Neuron, Ubongo…

Zowonjezereka, zosavuta kupanikizika, zidzasintha zotsatira zotsatizana

Pancreas, Chithokomiro…

Wolemera mu amkati michere, zimakhudza kupanga single selo kuyimitsidwa

Khoti Limodzi vs Selo Limodzi

Khungu limodzi

Selo limodzi

Ma cell opanda malire

Ma cell awiri: 10-40 μm

Zinthuzo zimatha kukhala minofu yowuma

Zinthuzo ziyenera kukhala minofu yatsopano

Kupanikizika kochepa kwa maselo oundana

Kuchiza kwa ma enzyme kungayambitse kupsinjika kwa ma cell

Palibe maselo ofiira a magazi omwe amafunika kuchotsedwa

Maselo ofiira amagazi amafunika kuchotsedwa

Nuclear imafotokoza bioinformation

Selo lonse limafotokoza bioinformation

Mafotokozedwe a Utumiki

Library

Njira yotsatirira

Voliyumu ya Data

Zitsanzo Zofunika

Minofu

10 × Genomics single-nuclei library

10x Genomics -Illumina PE150

100,000 amawerengedwa/maselo pafupifupi.100-200 Gb

Nambala ya cell: > 2 × 105

Cell conc.pa 700-1,200 cell/μL

≥ 200 mg

Kuti mumve zambiri zachitsogozo chokonzekera zitsanzo ndi kayendedwe ka ntchito, chonde omasuka kulankhula ndi a

Kuyenda kwa Ntchito Yantchito

Chitsanzo cha QC

Kuyesera kupanga

kutumiza chitsanzo

Kupereka zitsanzo

nyukiliya.kuyimitsidwa

Kudzipatula

Kukonzekera kwa Library

Kumanga laibulale

Kutsata

Kutsata

Kusanthula deta

Kusanthula deta

Pambuyo pogulitsa Services

Pambuyo pogulitsa ntchito


  • Zam'mbuyo:
  • Ena:

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    1.Spot clustering

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    2.Mawu a Marker kuchuluka komwe kumaphatikiza kutentha

    wps_doc_113.Maker kugawa majini m'magulu osiyanasiyana

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    4.Cell trajectory analysis/pseudotime

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