Ukulandelana Okude Okungafaki Amakhodi-Illumina
Izinzuzo Zesevisi
●Ukuhlaziywa okuhlangene kwe-mRNA kanye ne-lncRNANgokuhlanganisa ukulinganiswa kwemibhalo ye-mRNA nocwaningo lwe-lncRNA kanye nezinhloso zayo, kungenzeka ukuthola umbono ojulile wendlela yokulawula eyisisekelo sempendulo yamaseli.
●Ubuchwepheshe Obubanzi: Sicubungule amasampula angaphezu kuka-230,000, ahlanganisa izinhloso zamasampula ahlukahlukene kanye namaphrojekthi. Siletha ulwazi oluningi kuyo yonke iphrojekthi.
●Ukuhlaziywa okuhlangene kwe-mRNA kanye ne-lncRNA: Sihlanganisa ukulinganiswa kwemibhalo ye-mRNA nocwaningo lwe-lncRNA kanye nezinhloso zayo, okwenza kube nokwenzeka ukuthola umbono ojulile wendlela yokulawula eyisisekelo sempendulo yamaseli.
●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.
●Isichasiselo Esiphelele: Sisebenzisa izizindalwazi eziningi ukuze sichaze ngokusebenza i-Differentially Expressed Genes (DEGs) futhi senze ukuhlaziywa kokunotha okuhambisanayo. Le ndlela ebanzi inikeza ukuqonda ngezinqubo zamaseli kanye nama-molecule ezisekela impendulo ye-transcriptome, okuqinisekisa ukuthi uthola lonke ulwazi olungenzeka mayelana nedatha yokuhlolwa kwakho.
●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 | 10-16 Gb | Q30≥85% |
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 eluhlaza
- Idatha ye-QC
- Ukuqondanisa i-genome
- Isakhiwo sezakhi zofuzo (Ukuhlanganisa okunye, ukwenza ngcono isakhiwo sezakhi zofuzo kanye nokubikezela okusha kwezakhi zofuzo)
- Ukulinganiswa kokubonakaliswa kwezakhi zofuzo
- Ukuhlaziywa kokubonakaliswa kokwehluka
- Isichasiselo se-DEG kanye nokucebisa + Izakhi zofuzo eziqondiwe ze-lncRNA ezivezwe ngokuhlukile
- Ukuhlonza umbhalo oqoshiwe
- Ukuhlonza i-lncRNA (ukulondolozwa kwe-lncRNA kanye ne-lncRNA eyaziwayo)
- Ukubikezela izakhi zofuzo eziqondiwe ze-lncRNA
- Ukulinganiswa kokubonakaliswa kwe-lncRNA
- Ukuhlaziywa okuhlangene nedatha ye-mRNA
Ukuhlaziywa kwe-Differential Gene Expression (DEGs)
Ukulinganiswa kokubonakaliswa kwe-lncRNA - ukuhlanganiswa
Ukucebisa izakhi zofuzo ze-lncRNA
Ukuhlaziywa kwesimo se-mRNA kanye ne-lncRNA esihlangene – Isakhiwo se-Circos (isiyingi esiphakathi siyi-mRNA kanti i-inner cicrlce iyi-lncRNA)
Hlola intuthuko eyenziwe yizinsizakalo zokulandelana kwe-lncRNA ze-BMKGene ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
UJi, H. et al. (2020) 'Ukuhlonza, ukubikezela okusebenzayo, kanye nokuqinisekiswa okubalulekile kwe-lncRNA kwama-lncRNA ahlobene nokucindezeleka okubandayo esibindini samagundane',Imibiko Yesayensi2020 10:1, 10(1), amakhasi 1–14. doi: 10.1038/s41598-020-57451-7.
UJia, Z. et al. (2021) 'Ukuhlaziywa Kwe-Transcriptomic Okuhlanganisiwe Kwembula Indlela Yokuzivikela Yomzimba Yohlobo Lwe-Carp Oluhlala Njalo Lwe-CyHV-3',Imingcele ku-Immunology, 12, ikhasi. 687151. doi: 10.3389/FIMMU.2021.687151/BIBTEX.
UWang, XJ nabanye. (2022) 'Ukubekwa phambili okusekelwe ekuhlanganisweni kwe-Multi-Omics kwamanethiwekhi okulawula i-RNA ancintisanayo kumdlavuza wamaphaphu wamangqamuzana amancane: izici zama-molecular kanye nabantu abazosebenzisa izidakamizwa',Imingcele ku-Oncology, 12, ikhasi. 904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
UXiao, L. et al. (2020) 'Ukuhlukaniswa kwezakhi zofuzo kwenethiwekhi yokuxhumana kwezakhi zofuzo engaphansi kwe-photosynthesis ku-Populus',Ijenali ye-Plant Biotechnology, 18(4), amakhasi 1015–1026. doi: 10.1111/PBI.13270.
UZheng, H. et al. (2022) 'Inethiwekhi Yomhlaba Wonke Yokulawula Ukuvezwa Kwezakhi Zofuzo Okungahlelekile kanye Nokubonakaliswa Okungavamile Kwe-Metabolic Kumaseli Okuzivikela Emzimbeni Emvelweni Yezifo Zamathuna kanye ne-Hashimoto's Thyroiditis',Imingcele ku-Immunology, 13, ikhasi. 879824. doi: 10.3389/FIMMU.2022.879824/BIBTEX.
