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Usoro RNA otu nucleus

Ọganihu n'ime otu cell na usoro iwu ụlọ ọba akwụkwọ jikọtara ya na usoro ntinye dị elu na-enye ohere ọmụmụ mkpụrụ ndụ ihe nketa site na cell site na cell.Ọ na-eme ka nyocha nke sistemu dị omimi na nke zuru oke na ọnụ ọgụgụ sel dị mgbagwoju anya, nke ọ na-ezere nkpuchi nke ịdị iche iche ha site na iwere nkezi nke sel niile.

Otú ọ dị, ụfọdụ mkpụrụ ndụ adịghị mma ka e mee ya ka ọ bụrụ nkwusioru nke otu cell, ya mere, usoro nkwadebe ihe atụ ndị ọzọ - a na-ewepụta oghere site na anụ ahụ, ya bụ, a na-esi na anụ ahụ ma ọ bụ cell ewepụta ngwa ngwa ma kwadebe ya n'otu nkwusioru maka otu-nucleus. usoro cell.

BMK na-enye 10× Genomics ChromiumTM nke dabere na otu cell RNA ọrụ usoro.A na-eji ọrụ a eme ihe n'ọtụtụ ebe n'ọmụmụ ihe gbasara ọmụmụ ihe metụtara ọrịa, dị ka ọdịiche dị n'etiti cell immunodeficiency, ụdị ọrịa tumor, mmepe anụ ahụ, wdg.

Mgbawa transcriptome spatial: 10× Genomics

Platform: Illumina NovaSeq Platform


Nkọwa ọrụ

Bioinformatics

Nsonaazụ ngosi

Usoro nka

A na-enweta ikewapụ nke nuclei site na 10 × Genomics Chromium TM, nke nwere sistemụ microfluidics nke ọwa asatọ nwere ngafe abụọ.Na usoro a, a gel beads na barcodes na primer, enzymes na otu nucleus na-etinyere na mmanụ mmanụ nanoliter, na-emepụta Gel Bead-in-Emulsion (GEM).Ozugbo e guzobere GEM, a na-eme lysis cell na ntọhapụ nke barcode na GEM ọ bụla.A na-atụgharị mRNA n'ime mkpụrụ ndụ cDNA nwere koodu barcode 10 × na UMI, ndị na-aga n'ihu maka iwu ụlọ ọba akwụkwọ nhazi usoro.

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Anụ ahụ adịghị mma maka nkwadebe nkwusioru otu cell

Cell / anụ ahụ

Ihe kpatara ya

Megharịa anụ ahụ jụrụ oyi

Enweghị ike ịnweta ụlọ ọrụ ọhụrụ ma ọ bụ ndị echekwara ogologo oge

Mkpụrụ ndụ akwara, Megakaryocyte, abụba…

Dayameta cell dị oke ibu ịbanye ngwa

Imeju…

Ọ na-esighi ike ịgbaji, enweghị ike ịmata otu mkpụrụ ndụ

Mkpụrụ ndụ Neuron, Brain…

Ọ dị nro karịa, dị mfe nrụgide, ga-agbanwe nsonaazụ usoro

Pancreas, thyroid…

Ọgaranya na enzymes endogenous, na-emetụta mmepụta nke nkwusioru otu cell

Otu-nucleus vs Single-cell

Otu-nucleus

Otu cell

Dayameta cell na-akparaghị ókè

Dayameta cell: 10-40 μm

Ihe nwere ike ịbụ anụ ahụ jụrụ oyi

Ihe ga-abụrịrị anụ ahụ dị ọhụrụ

Obere nchekasị nke mkpụrụ ndụ oyi kpọnwụrụ

Ọgwụgwọ enzyme nwere ike ibute mmeghachi omume nrụgide cell

Ọ dịghị mkpụrụ ndụ ọbara uhie ka a ga-ewepụ

Ọ dị mkpa iwepụ mkpụrụ ndụ ọbara uhie

Nuklia na-egosipụta ozi bioin

Mkpụrụ ndụ niile na-egosipụta ozi bioin

Nkọwapụta ọrụ

Ụlọ akwụkwọ

Usoro usoro

Olu data

Ihe atụ chọrọ

Anụ ahụ

10× Genomics otu-nuclei ọba akwụkwọ

10x Genomics -Ilumina PE150

100,000 na-agụ / cell ihe ruru.100-200 Gb

Nọmba ekwentị:>2× 105

Cell conc.na 700-1,200 cell / μL

≥ 200 mg

Maka nkọwa ndị ọzọ na ntụzịaka nkwadebe nlele na usoro ọrụ ọrụ, biko nweere onwe gị ịgwa a

Usoro ọrụ ọrụ

Ihe atụ QC

Nnwale imewe

nnyefe sample

Nbufe ihe atụ

nuclear.nkwusioru

Mwepu nke nuklia

Nkwadebe ụlọ akwụkwọ

Ịrụ ụlọ akwụkwọ

Usoro

Usoro

Nyocha data

Nyocha data

Mgbe ire Ọrụ

Mgbe-ire ọrụ


  • Nke gara aga:
  • Osote:

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    1.Spot ụyọkọ

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    2.Marker okwu nke ukwuu clustering heatmap

    wps_doc_113.Maker nkesa nkesa na ụyọkọ dị iche iche

    wps_doc_12

    4.Cell trajectory analysis/pseudotime

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