Ukulandelana kwe-CircRNA-Illumina
Izici
● Ukuncipha kwe-rRNA kulandelwa ukulungiswa komtapo wolwazi oqondisa izinkomba, okuvumela idatha yokulandelana kwemigqa ethile.
● Ukuhamba komsebenzi kwe-Bioinformatic kwenza ukubikezela kwe-circRNA kanye nokulinganisa ukubonakaliswa.
Izinzuzo Zesevisi
●Ubuchwepheshe Obubanzi: Sicubungule amasampula angaphezu kuka-20,000, ahlanganisa izinhlobo ezahlukene zamasampula, siletha ulwazi oluningi kuyo yonke iphrojekthi.
●Ukulawulwa Kwekhwalithi Okuqinile: Sisebenzisa amaphuzu okulawula ayisisekelo kuzo zonke izigaba, kusukela ekulungiseleleni amasampula kuya ekulungiseleleni umtapo wolwazi, ukulandelana kanye ne-bioinformatics. Ukuqapha kwethu ngokucophelela kuqinisekisa ukulethwa kwemiphumela esezingeni eliphezulu njalo.
●Imitapo Yezincwadi Ye-RNA Ebanzi Kakhulu:Sisebenzisa ukuncipha kwe-rRNA esikhundleni sokuncipha kwe-RNA eqondile ekulungiseleleni kwethu kwangaphambi komtapo wolwazi, siqinisekisa ukuthi idatha yokulandelana ayihlanganisi i-circRNA kuphela kodwa ne-mRNA kanye ne-lncRNA, okuvumela ukuhlaziywa okuhlangene kwalawa masethi edatha.
●Ukuhlaziywa Okuzikhethela Kwamanethiwekhi E-Endogenous RNA (ceRNA) Okuncintisanayo: Ukunikeza ukuqonda okujulile ngezindlela zokulawula amaseli.
● Usekelo Lwangemva Kokuthengisa: Siyaqonda ukuthi kubaluleke kangakanani ukuba khona, yingakho ukuzibophezela kwethu kudlulela ngale kokuphothulwa kwephrojekthi ngesikhathi senkonzo yezinyanga ezintathu ngemuva kokuthengisa. Phakathi nalesi sikhathi, sinikeza ukulandelwa kwephrojekthi, usizo lokuxazulula izinkinga, kanye nezikhathi ze-Q&A 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 | 16-20 Gb | Q30≥85% |
Izidingo Zesampula:
Ama-Nucleotide:
| I-Conc.(ng/μl) | Inani (μg) | Ubumsulwa | Ubuqotho |
| ≥ 80 | ≥ 0.8 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Ukungcoliswa kwephrotheni noma i-DNA okulinganiselwe noma okungekho nhlobo kukhonjisiwe kujeli. | I-RIN≥6.0; 5.0≥28S/18S≥1.0; ukuphakama okulinganiselwe noma okungekho kwesisekelo |
● Izitshalo:
Impande, Isiqu noma Ipetali: 450 mg
Iqabunga noma Imbewu: 300 mg
Izithelo: 1.2 g
● Isilwane:
Inhliziyo noma Amathumbu: 450 mg
I-Viscera noma Ubuchopho: 240 mg
Imisipha: 600 mg
Amathambo, Izinwele noma Isikhumba: 1.5g
● Ama-arthropod:
Izinambuzane: 9g
I-Crustacea: 450 mg
● Igazi eliphelele:Amapayipi ama-2
● Amaseli: 106 amaseli
● I-Serum ne-Plasma: 6 ml
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.
Ukugeleza Komsebenzi Wesevisi
Umklamo wokuhlola
Ukulethwa kwesampula
Ukukhishwa kwe-RNA
Ukwakhiwa komtapo wolwazi
Ukulandelana
Ukuhlaziywa kwedatha
Izinsizakalo zangemuva kokuthengisa
I-Bioinformatics
- Idatha ye-QC
- Ukuqondanisa i-genome
- Ukuhlonza i-CircRNA
- Ukubonakaliswa kwe-CircRNA
- Ukuhlaziywa kokubonakaliswa kokwehluka
Ukubikezela kwe-circrRNA: ukusatshalaliswa kwe-chromosome
Ama-circrRNA ahlukaniswe ngokuhlukile – i-volcano plot
Ama-circrRNA ahlukaniswe ngokuhlukile - ukuhlanganiswa kwe-hierarchical
Ukucebisa okusebenzayo kwezakhi zofuzo ze-circRNA
Hlola intuthuko yocwaningo eyenziwe yizinsizakalo zokulandelana kwe-BMKGene' circRNA ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
UWang, X. et al. (2021) 'I-CPSF4 ilawula ukwakheka kwe-circRNA kanye nokuthuliswa kwezakhi zofuzo eziqondiswa yi-microRNA ku-carcinoma ye-hepatocellular',I-Oncogene2021 40:25, 40(25), amakhasi 4338–4351. doi: 10.1038/s41388-021-01867-6.
UXia, K. et al. (2023) 'I-transcriptome ye-X oo-responsive yembula indima ye-RNA133 eyindilinga ekumelaneni nezifo ngokulawula ukubonakaliswa kwe-OsARAB elayisini',Ucwaningo lwe-Phytopathology, 5(1), amakhasi 1–14. doi: 10.1186/S42483-023-00188-8/FIGURES/6.
Y, H. et al. (2023) 'I-CPSF3 ilungisa ibhalansi yemibhalo eyindilinga neqondile ku-carcinoma yesibindi'. doi: 10.21203/RS.3.RS-2418311/V1.
UZhang, Y. et al. (2023) 'Ukuhlolwa okuphelele kwama-circRNA ku-cardiomyopathy ye-cirrhotic ngaphambi nangemva kokufakelwa kwesibindi',I-Immunopharmacology Yomhlaba Wonke, 114, ikhasi. 109495. doi: 10.1016/J.INTIMP.2022.109495.
