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Chingwe cha Nina

Kukula kwa njira zomangirira limodzi ndi chizolowezi, kuphatikiza ndi mawonekedwe apamwamba kwambiri, amasinthana majini a gene. Kuthetsa kumeneku kumathandizanso kuyang'ana kwambiri kwa anthu ambiri, kuthana ndi malire omwe amagwirizana ndi mitundu yosokoneza pakati pa maselo onse ndikusunga zowona zenizeni mkati mwa anthu awa. Ngakhale Sex-Cell RNA (SPRNA-SEQ) ali ndi zovuta zingapo, zimakumana ndi ziwalo zina pomwe zomwe zimapangidwa ndi minofu imodzi zimakhala zovuta ndipo zimafunikira zitsanzo zatsopano. Ku BMKGene, timathana ndi vutoli popereka zingwe zosakwatiwa zamimba (SNRNA-Seq) pogwiritsa ntchito boma la 10x genomics chromium. Njirayi imakulitsa mawonekedwe a zitsanzo zowoneka bwino kuwunika pamakina a cell imodzi.

Kupatula kwa nuclei kumachitika kudzera mu encricy genomics cromium chip, chopangidwa ndi microfluidics eyiti ndi zopinga ziwiri. M'dongosolo lino, mikanda ya gel yophatikiza mabizinesi, primers, ma enzyme, ndipo ma elzy amodzi omwe amayang'aniridwa m'mafuta a Nanoliter-emulsion (gel). Kutsatira mapangidwe a Gem, cell lysis ndi mabatani amapezeka mkati mwa gemu iliyonse. Pambuyo pake, mamolekyulu a MRNA amapezekanso ku CDNAS, kuphatikiza ma barcode 10x komanso zidziwitso zapadera zodziwika bwino (Umis). A CDNAS awa akugonjera zomangamanga zowoneka bwino, kuwongolera kufufuza kokhazikika komanso kokwanira kwa majini a generani pamlingo umodzi.

Pulatifomu: 10 × Genomical Chromium ndi Suldina Platifomu

Lumikizanani nafe

Zambiri

Bioinfoctic

Zotsatira za Demo

Zolemba zolembedwa

CHENJEZO

Kupatula kwa nuclei kumachitika ndi 10 × geromics chromium ™, yomwe ili ndi microfludiidics eyiti ndi zopinga ziwiri. M'dongosolo lino, mikanda ya gel ya gel yokhala ndi ma barcode ndi primer, ma enzyme ndi zigawo zingapo zimakhazikika mu dontho lamafuta a Nanoliter, kupanga gelsion-emulsion (rince). Kamodzi gemu atapangidwa, cell lysis ndi kumasulidwa kwa mabizinesi amachitidwa mu gemu iliyonse. A MRNA amasinthidwa kukhala ma molekyulu a CDna okhala ndi 10 × ma barcodes ndi umi, zomwe zimakhudzidwanso ndi ntchito yomanga library yolowera.

Mawonekedwe

● Kukonzekera kwa Emecle-Eclei kuchokera ku zitsamba zazifupi

● Kupanga gel bead-in-emulsion (Gem) kutsatiridwa ndi CDna Synthesis

● Mbale aliyense mu miyala yamtengo wapatali yomwe imadzaza ndi magawo anayi:

Poly (dt) mchira wa MRNA yoyambira ndi CDna Synthesis,

Chidziwitso chapadera chazomwe chimakhala (UMI) kuti mukonze tsankho

10x Barcode

Kumangiriza Kuwerenga Kwakanthawi 1

Ubwino

Zithunzi zosakwatiwa za roquus RNA zimayendetsa malire a seva imodzi ya RAN, zomwe zimakuthandizani:

● Kugwiritsa ntchito zitsanzo za zitsanzo zopanda zitsanzo osati zochepa chabe za zitsanzo zatsopano

● Maselo ochepa maselo ozizira poyerekeza ndi enzymatic mankhwala atsopano maselo atsopano, omwe amawonetsedwa mu deta yosinthira mu mawonekedwe a majini ocheperako

● Palibe chifukwa chochotsera maselo ofiira a magazi

● Muliko osavomerezeka

● Zitsanzo zambiri zomwe ndizoyenera kusanthula, kuphatikiza mitundu yovuta komanso yosalimba ya minofu yomwe imakonda kupindika kapena chiwonongeko pakasule minofu

Zitsanzo zomwe sizingasanthuledwa ndi sex-cell rna squening ndipo ndizoyenera kubereka kwa Nuclei RNA:

Khungu / minolu	Chifukwa
Zosasinthika	Takanika kupeza mabungwe atsopano kapena osungika
Minofu cell, megakavucyte, mafuta ...	Mulingo wa cell ndi waukulu kwambiri kuti ulowetse chida
Chiwindi ...	Osalimba kwambiri kuti athyole, osatha kusiyanitsa maselo amodzi
Neuron cell, ubongo ...	Omvera kwambiri, osavuta kupsinjika, asintha zotsatira zoyeserera
Pancreas, Chithokomiro ...	Olemera mu michere ya Eyals, akukhudza kupanga kwa ma cell

Nyuzi imodzi vs imodzi

Osakwatiwa	Cell-cell
Mulingo wopanda malire	Mulingo wa maselo: 10-40 μm
Zinthuzi zitha kukhala zoundana	Zinthuzo ziyenera kukhala zokhala ndi minofu yatsopano
Kupsinjika kwa maselo ocheperako	Mankhwala a enzyme amatha kuyambitsa mavuto a cell
Palibe maselo ofiira amwazi omwe amafunikira kuchotsedwa	Maselo ofiira amafunika kuchotsedwa
Nuclear Ikufotokoza Bioinformation	Kholo lonse limafotokoza bioinfortion

Kulembana

Zofunikira Zitsanzo

Nkhokwe ya mabuku

Njira Yolowetsa

Zambiri Zotsimikiziridwa

Kuwongolera kwapadera

Nyama yanyama ≥ 200 mg

Chomera minofu ≥ 400 mg

10x Genomics Snomics CDNE CDNA

Audina Pe150

100K pe imawerenga pa foni

(100-200 GB)

700-1200 nuclei / μl ndi kukhulupirika kwa nuclei kumachitika pansi pa microscope

Kuti mumve zambiri pa zitsanzo zokonzekera ndi ntchito yogwira ntchito, chonde khalani omasuka kuyankhula ndi aKatswiri wa BMKGEne

Ntchito Yogwira Ntchito

M'mbuyomu: Chomera / chinyama chonse

Ena: Kusanthula Kwambiri

Mulinso kuwunika:

● Kuwongolera kwapadera: Chiwerengero cha maselo, chizindikiritso, chizindikiritso cholondola cha maselo, RNA ma molekyulu ndi mawu owerengera

● Kusanthula kwamkati:

Cell osamangitsidwa ndi tsatanetsatane

Kusanthula kosiyanasiyana: Kuzindikira mabatani

Zogwirira ntchito ndi zopindulitsa za tsambali

● Kusanthula kwa Gulu:

Kuphatikiza kwa deta

Kusanthula kosiyanasiyana: Kuzindikira magikitsi

Zogwirira ntchito ndi zopindulitsa madigiri

● Kusanthula ophunzira:

Kusanthula kwa cell

Kusanthula kwa Pseudotime

Kusanthula kwa maselo (celcondedb)

RACE ikhazikike kuwunika (GSEA)

Kusanthula kwamkati

Kusankhidwa kwa cell:

Kusanthula kosiyanasiyana: Tsitsani dinesi

Kusanthula kwa magulu

Kusanthula kosiyanasiyana: Madigiri

Kusanthula kwapamwamba:

Kusanthula kwa Pseudotime:

Kusanthula kwa cell:

Funsani zomwe zikugwirizana ndi ntchito za BMKGene za BMNSGey RNA zigawo za RNA Chromium mu 10x chromium mu zofalitsa izi:

Wang, L. et al. (2021) 'Kusanthula kwapaderaZochitika za National Academy of Sayansi ya United States of America, 118 (2), p. E20055590118. Doi: 10.1073 / PNAS.2005590118

Zheng, H. et al. "Mabanja mu Immunology, 13, p. 879824. Doi: 10.3389 / FIMMU.2022.879824 / BIBEREX.

Tian, H. et al. (2023) 'Kutanthauzira kwa Cell Syolg sikukuvulirani kukhazikika kwa ma leukocyte pambuyo pa katemera ndi Edctocdwand Eddatla Tarda Boundda Inrda ku Flonder (Paralichthys Olivaces)',Ufale, 566, p. 739238. Doi: 1016016 / J.Aacal.2023.739238.

Yu, y. et al. .Khansa yam'mimba, 26 (5), mas. 798-813. Doi: 10.1007 / S10120-023-01409-x / menics.