Imikhiqizo

I-Exosomal mRNA/LncRNA/CirrcRNA Sequencing-Illumina

Isithombe Esifakiwe se-mRNA/LncRNA/CirrcRNA Sequencing-Illumina

Ama-Exosome angama-vesicle amancane akhishwa amaseli, ngokuvamile asukela kuma-nanometer angu-30 kuya kwangu-100 ububanzi. Lawa ma-vesicle aqukethe i-RNA ehlukahlukene. Ama-Exosome kukholakala ukuthi adlala indima ebalulekile ekuxhumaneni phakathi kwamangqamuzana, izimpendulo zomzimba, kanye nokuthuthukiswa kwezifo, futhi angasakazwa kwezinye izingxenye zomzimba ngoketshezi lomzimba njenge-plasma, amathe, kanye nomchamo. Aphethe ama-biomolecule athile ukulawula imisebenzi yamaseli amukelayo, ethonya izimo zomzimba zamaseli. Ama-Exosome nawo abhekwa njengadlala indima ebalulekile ekuthuthukisweni kwezifo, okuhlanganisa umdlavuza, izifo eziwohlokayo zemizwa, kanye nezimo zokuvuvukala. Ucwaningo ngama-exosome lunikeza ukuqonda okusha kanye nezindlela zokuxilonga, ukwelashwa, kanye nokuvimbela izifo.

Xhumana nathi

Imininingwane Yesevisi

I-Bioinformatics

Imiphumela Yedemo

Izici

● Kusetshenziswa ama-probe akhethekile ukuqeda i-ribosomal cDNA.

● Ukwakhiwa komtapo wolwazi oqondene ne-strand.

● Umsebenzi owodwa wokuthola idatha ye-mRNA, i-lncRNA kanye ne-circRNA ngasikhathi sinye.

Izinzuzo Zesevisi

● Ukulawulwa Kwekhwalithi Okuqinile: Sisebenzisa amaphuzu okulawula ayisisekelo kuzo zonke izigaba, kusukela ekulungiseleleni amasampula kanye nomtapo wolwazi kuya ekulandeleni nasekubhaleni ulwazi nge-bioinformatics. Lokhu kuqapha ngokucophelela kuqinisekisa ukulethwa kwemiphumela esezingeni eliphezulu njalo.

● Ukusekelwa Ngemva Kokuthengisa: Ukuzibophezela kwethu kudlulela ngale kokuqedwa kwephrojekthi ngesikhathi sesevisi yezinyanga ezintathu ngemuva kokuthengisa. Phakathi nalesi sikhathi, sinikeza ukulandelwa kwephrojekthi, usizo lokuxazulula izinkinga, kanye nezikhathi zemibuzo nezimpendulo ukuze kuxazululwe noma yimiphi imibuzo ehlobene nemiphumela.

Izidingo Zesampula Nokulethwa

Umtapo Wolwazi	Ipulatifomu	Idatha enconyiwe	Idatha ye-QC
umtapo wolwazi oqondisiwe we-rRNA onciphile	I-Illumina PE150	10-16 Gb	Q30≥85%

Izidingo Zesampula

Ama-Nucleotide:

Inani (ng)	Ivolumu (μL)
40	20

Izicubu:

I-Exosome: 100 μL; 40 μg; >10^9 izinhlayiya/mL
I-Serum/plasma: 5 mL
Isithako esingaphezu kwesijwayelekile se-cell culture: 100 mL
Umchamo: 100 mL
Amathe: 15 mL
Ukuphuma kwempumulo: 15 mL
Uketshezi lwe-cerebrospinal: 15 mL
Uketshezi lwe-Amniotic: 50 mL
Isidoda: 5 mL
Inyongo: 5 mL
Ama-Ascites: 50 mL
Ubisi: 15 mL
Uketshezi lwe-Follicular: 15 mL
I-supernatant yendle: 50 mL
Isithako esikhulu se-microbial culture: 100 mL
Izicubu zezilwane: 3 g
Izicubu zesitshalo: 1 g

Ukulethwa Kwesampula Okunconyiwe

Isitsha: ishubhu le-centrifuge elingu-2 mL (Akukhuthazwa ukusebenzisa i-tin foil)
Ukulebula kwesampula: Iqembu + ukukopisha isib. A1, A2, A3; B1, B2, B3.
Ukuthunyelwa:
1. Iqhwa elomile: Amasampula kudingeka apakishwe ezikhwameni bese egqitshwa eqhweni elomile.
2. Amashubhu e-RNAstable: Amasampula e-RNA angomiswa eshubhu lokuzinzisa i-RNA (isb. i-RNAstable®) bese ethunyelwa ekushiseni kwegumbi.