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

Lumikizanani nafe

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.

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