i-circRNA sequencing-Illumina
Izici
● Ukuncipha kwe-rRNA okulandelwa ukulungiswa komtapo wolwazi, okuvumela ukulandelana kwedatha eqondene ngqo ne-strand.
● Ukugeleza komsebenzi kwe-Bioinformamatic kunika amandla ukubikezela kwe-circRNA kanye nokulinganisa kwenkulumo
Izinzuzo Zesevisi
●Imitapo yolwazi ye-RNA ebanzi:sisebenzisa ukuncipha kwe-rRNA esikhundleni sokuncishiswa kwe-RNA ngomugqa ekulungiseleleni kwethu kwangaphambi kwelabhulali, siqinisekisa ukuthi idatha yokulandelana ayihlanganisi i-circRNA kuphela kodwa futhi i-mRNA kanye ne-lncRNA, okuvumela ukuhlaziywa okuhlanganyelwe kulawa madathasethi.
●Ukuhlaziya ongakukhetha kwamanethiwekhi okuncintisana e-endogenous RNA (ceRNA).: ukuhlinzeka ngemininingwane ejulile ezindleleni zokulawula amaselula
●Ubungcweti Olubanzi: Ngomlando wokucubungula amasampula angaphezu kuka-20,000 kwa-BMK, ahlanganisa izinhlobo zamasampula ahlukahlukene namaphrojekthi e-lncRNA, ithimba lethu liletha ingcebo yesipiliyoni kuyo yonke iphrojekthi.
●Ukulawulwa Kwekhwalithi Okuqinile: sisebenzisa amaphuzu okulawula abalulekile kuzo zonke izigaba, kusukela kusampula nokulungiselela umtapo wezincwadi ukuya ekulandelaneni kanye ne-bioinformatics.Lokhu kuqapha ngokucophelela kuqinisekisa ukulethwa kwemiphumela ehlala njalo yekhwalithi ephezulu.
● Ukusekelwa Kwangemva Kokuthengisa: Ukuzinikela kwethu kudlulela ngalé kokuphothulwa kwephrojekthi ngenkathi yesevisi yezinyanga ezi-3 ngemva kokuthengisa.Ngalesi sikhathi, sinikezela ngokulandelelwa kwephrojekthi, usizo lokuxazulula inkinga, namaseshini e-Q&A ukuze siphendule noma yimiphi imibuzo ehlobene nemiphumela.
Izidingo Zesampula Nokulethwa
| Umtapowolwazi | Inkundla | Idatha enconyiwe | Idatha ye-QC |
| I-Poly A inothisiwe | I-Ilumina PE150 | 16-20 Gb | Q30≥85% |
Izimfuneko zesampula:
Ama-Nucleotides:
| I-Conc.(ng/μl) | Inani (μg) | Ubumsulwa | Ubuqotho |
| ≥ 100 | ≥ 0.5 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Amaprotheni anomkhawulo noma awekho nhlobo noma ukungcola kwe-DNA okuboniswa kujeli. | Okwezitshalo: RIN≥6.5; Okwezilwane: RIN≥7.0; 5.0≥28S/18S≥1.0; ukuphakama okulinganiselwe noma okungenakho |
● Izitshalo:
Izimpande, Isiqu noma I-Petal: 450 mg
Iqabunga noma Imbewu: 300 mg
Isithelo: 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-arthropods:
Izinambuzane: 9g
I-crustacea: 450 mg
● Igazi eliphelele:2 amashubhu
● Amaseli: 106 amaseli
● I-Serum ne-Plasmaawu :6ml
Ukulethwa Kwesampula Okunconyiwe
Isitsha: 2 ml centrifuge tube (Tin foil ayinconywa)
Ukulebula okuyisampula: Ukuphindaphinda kweqembu+isb A1, A2, A3;B1, B2, B3....
Ukuthunyelwa:
1. Iqhwa elomile: Amasampula adinga ukupakishwa ezikhwameni futhi agqitshwe eqhweni elomile.
2. Amashubhu e-RNAstable: Amasampula e-RNA angomiswa ngeshubhu lokuzinzisa le-RNA(isb. i-RNAstable®) futhi athunyelwe ekamelweni lokushisa.
Ukugeleza Komsebenzi Wesevisi
Idizayini yokuhlola
Ukulethwa kwesampula
Ukukhishwa kwe-RNA
Ukwakhiwa komtapo wolwazi
Ukulandelanisa
Ukuhlaziywa kwedatha
Amasevisi angemuva kokuthengisa
I-Bioinformatics
ukubikezela kwe-circRNA: ukusatshalaliswa kwe-chromosomal
Ama-circRNA Avezwe Ngokuhlukile - Isakhiwo sentaba-mlilo
Ama-circRNA Avezwe Ngokuhlukile – ukuhlanganisa okulandelanayo
Ukucebisa okusebenzayo kwezakhi zofuzo zomsingathi we-circRNA
Hlola intuthuko yocwaningo eqhutshwa izinsiza zokulandelana ze-BMKGene' circRNA ngeqoqo elikhethiwe lokushicilelwe.
Wang, X. et al.(2021) 'I-CPSF4 ilawula ukwakheka kwe-circRNA kanye ne-microRNA mediated gene silenceng ku-hepatocellular carcinoma', i-Oncogene 2021 40:25, 40(25), amakhasi 4338–4351.doi: 10.1038/s41388-021-01867-6.
Xia, K. et al.(2023) I-'X oo-responsive transcriptome iveza indima ye-RNA133 eyindilinga ekumelaneni nezifo ngokulawula ukuvezwa kwe-OsARAB elayisini', Phytopathology Research, 5(1), amakhasi 1-14.doi: 10.1186/S42483-023-00188-8/AMAFIGURES/6.
Y, H. et al.(2023) 'I-CPSF3 ilungisa ibhalansi yemibhalo eyisiyingi neyomugqa ku-hepatocellular carcinoma'.doi: 10.21203/RS.3.RS-2418311/V1.
U-Zhang, Y. et al.(2023) 'Ukuhlolwa okuphelele kwe-circRNAs ku-cirrhotic cardiomyopathy ngaphambi nangemva kokufakelwa kwesibindi', International Immunopharmacology, 114, p.109495. doi: 10.1016/J.INTIMP.2022.109495.
