I-Eukaryotic mRNA Sequencing-NGS
Izici
● Ukuthwebula i-poly mRNA ngaphambi kokulungiswa komtapo wolwazi
● Ukulungiswa komtapo wezincwadi we-mRNA ongakhetha kukho ukuze unike amandla ukuthola idatha yokulandelana kwe-strand ethize
● Ukuhlaziywa kwe-Bioinformamatic ye-gene expression kanye ne-transcript structure
Izinzuzo
●Ubungcweti Olubanzi: Ithimba lethu licubungule amasampula angaphezu kuka-600,000 kwa-BMKgene, ahlanganisa izinhlobo zamasampula ahlukahlukene njengamasiko amaseli, izicubu, noketshezi lomzimba. Siletha ingcebo yolwazi kuwo wonke amaphrojekthi futhi siqede ngempumelelo amaphrojekthi angaphezu kuka-100,000 mRNA-Seq ezizindeni ezihlukahlukene zocwaningo.
●Ukulawulwa Kwekhwalithi Okuqinile: Sisebenzisa amaphuzu okulawula abalulekile kuzo zonke izigaba, kusukela kusampula nokulungiselela umtapo wezincwadi ukuya ekulandeleni nasekulandeleni i-bioinformatics. Lokhu kuqapha ngokucophelela kuqinisekisa ukulethwa kwemiphumela ehlala njalo yekhwalithi ephezulu, kukuqinisekisa ukuthi iphrojekthi yakho isezandleni ezithembekile.
●Isichasiselo Esibanzi: Sisebenzisa imininingo egciniwe eminingi ukuze sichaze ngokusebenza Izakhi Zofuzo Ezivezwe Ngokuhlukile (DEGs) futhi senze ukuhlaziya okunothisayo okuhambisanayo. Le ndlela ebanzi ihlinzeka ngemininingwane ngezinqubo zamangqamuzana namangqamuzana angaphansi kwempendulo ye-transcriptome, iqinisekisa ukuthi unolwazi olugcwele ngemiphumela yephrojekthi yakho.
●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 | Isu lokulandelanisa | Idatha iyanconywa | Ikhwalithi yokulawula |
| I-Poly A inothisiwe | I-Ilumina PE150 I-DNBSEQ-T7 | 6 -10 Gb | Q30≥85% |
Izimfuneko zesampula:
Ama-Nucleotides:
| Conc.(ng/μl) | Inani (μg) | Ubumsulwa | Ubuqotho | |
| Ilabhulali Ejwayelekile | ≥ 10 | ≥ 0.2 | 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≥4.0; Okwezilwane: RIN≥4.5; 5.0≥28S/18S≥1.0; ukuphakama okulinganiselwe noma okungenakho |
| Ilabhulali yokuqondisa | ≥ 10 | ≥ 0.2 | 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≥4.0; Okwezilwane: RIN≥4.5; 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: 300 mg
I-Viscera noma Ubuchopho: 240 mg
Imisipha: 450 mg
Amathambo, Izinwele noma Isikhumba: 1g
● Ama-arthropods:
Izinambuzane: 6g
I-crustacea: 300 mg
● Igazi lonke: 1 ithubhu
● Amaseli: 106 amaseli
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 ethebula e-RNAs: 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
I-Eukaryotic I-mRNA ukulandelana kokuhlaziywa komsebenzi
I-Bioinformatics
ØUkulawulwa kwekhwalithi yedatha eluhlaza
ØUkuqondanisa kofuzo lwenkomba
ØUkuhlaziywa kwesakhiwo sombhalo
ØUbuningi bokuveza
ØUkuhlaziywa kwesisho esihlukile
ØIsichasiselo somsebenzi nokunothisa
Reference Genome Alignment
Ukugcwaliswa Kwedatha
Ukuhlanganiswa kwesampula nokuhlolwa kokuphindaphindeka kwezinto eziphilayo
Izakhi zofuzo ezivezwe ngokuhlukile (DEGs)
Isichasiselo esisebenzayo sama-DEGs
Ukuthuthukisa Okusebenzayo kwama-DEGs
Hlola intuthuko egqugquzelwa izinsiza zokulandelana ze-Eukaryotic NGS mRNA ze-BMKGene ngeqoqo elikhethiwe lokushicilelwe.
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Jin, K. et al. (2022) 'Izici Zokubhala ze-TCP Ezihilelekile Ekuthuthukisweni Kwe-Shoot ye-Ma Bamboo (Dendrocalamus latiflorus Munro)', Frontiers in Plant Science, 13, p. 884443. doi: 10.3389/FPLS.2022.884443/BIBTEX.
Wen, X. et al. (2022) 'I-Chrysanthemum lavandulifolium genome kanye nendlela yamangqamuzana engaphansi kwezinhlobo ezihlukahlukene ze-capitulum', Ucwaningo Lwezokulima, 9. doi: 10.1093/HR/UHAB022.
U-Zhang, u-Yujie et al. (2023) 'I-cascade nanoreactor yokuthuthukisa ukwelashwa kwe-sonodynamic kumdlavuza we-colorectal nge-synergistic ROS augment kanye nokuvinjelwa kwe-autophagy', Nano Namuhla, 49, p. 101798. doi: 10.1016/J.NANTOD.2023.101798.
