Ukulandelana kwe-Prokaryotic RNA
Izici
● Ukucubungula isampula ye-RNA kuhilela ukuncipha kwe-rRNA kulandelwe ukulungiswa komtapo wezincwadi we-RNA oqondisiwe.
● Ukuhlaziywa kwe-Bioinformatic okusekelwe ekuqondeni i-genome yokubhekisela
● Ukuhlaziywa kufaka phakathi ukubonakaliswa kwezakhi zofuzo kanye nama-DEG kodwa futhi nesakhiwo sombhalo kanye nokuhlaziywa kwe-sRNA
Izinzuzo Zesevisi
●Ukulawulwa Kwekhwalithi Okuqinile: sisebenzisa amaphuzu okulawula ayisisekelo kuzo zonke izigaba, kusukela ekulungiseleleni isampula kanye nomtapo wolwazi kuya ekulandeleni nasekubhaleni ulwazi nge-bioinformatics. Lokhu kuqapha ngokucophelela kuqinisekisa ukulethwa kwemiphumela esezingeni eliphezulu njalo.
●Idatha Yokulandelana Eqondile Ye-Strand: ngenxa yokuthi ukulungiswa komtapo wezincwadi we-RNA kuyaqondisa, okuvumela ukuhlonza imibhalo ephikisana nemizwa.
●Ukuhlaziywa Okuphelele Okuqondiswe Kuma-Transcriptomes e-Prokaryotic: ipayipi le-bioinformatic alihlanganisi nje kuphela ukuhlaziywa kokubonakaliswa kwezakhi zofuzo kodwa futhi nokuhlaziywa kwesakhiwo sombhalo, okuhlanganisa ukuhlonza ama-operons, ama-UTR kanye nama-promoters. Lihlanganisa nokuhlaziywa kwama-sRNA, okungukuthi isichasiselo kanye nokubikezela kwesakhiwo sesibili kanye nezinhloso.
●Usekelo Lwangemva Kokuthengisa: ukuzibophezela kwethu kudlulela ngale kokuqedwa kwephrojekthi ngesikhathi sesevisi 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 | Isu lokulandelana kwezigaba | Kunconywa idatha | Ikhwalithi yokulawula |
| umtapo wolwazi oqondisiwe we-rRNA onciphile | I-Illumina PE150 | 1-2 Gb | Q30≥85% |
Izidingo Zesampula:
| I-Conc.(ng/μl) | Inani (μg) | Ubumsulwa | Ubuqotho |
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Ukungcoliswa kwephrotheni noma i-DNA okulinganiselwe noma okungekho nhlobo kukhonjisiwe kujeli. | I-RIN≥6.5 |
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
Ukulethwa kwesampula
Ukwakhiwa komtapo wolwazi
Ukulandelana
Ukuhlaziywa kwedatha
Izinsizakalo zangemuva kokuthengisa
Ukuhamba Komsebenzi Wokuhlaziya I-Bioinformatic
Kufaka phakathi Ukuhlaziywa Okulandelayo:
● Ukulawulwa kwekhwalithi yedatha eluhlaza
● Ukuqondanisa ne-genome yokubhekisela
● Ukuhlolwa Kwekhwalithi Yomtapo Wolwazi: Ukungahleliwe kokuqhekeka kwe-RNA, usayizi wokufaka kanye nokugcwala kokulandelana
● Isichasiselo esisebenzayo sezakhi zofuzo zokubhala ikhodi ezibikezelwe
● Ukuhlaziywa Kwezincazelo: Ukuhlaziywa Kokuhlobana kanye Nokuhlaziywa Kwezingxenye Eziyinhloko (i-PCA)
● Ukuvezwa Kwezakhi Zofuzo Ezihlukile (ama-DEG)
● Isichasiselo esisebenzayo kanye nokucebisa ama-DEG
● Ukuhlaziywa kwe-sRNA: ukubikezela, isichasiselo, inhloso, kanye nokubikezela kwesakhiwo sesibili
● Ukuhlaziywa Kwesakhiwo Sombhalo: ama-operon, izikhundla zokuqala nezokugcina, isifunda esingahunyushwanga (i-UTS), iphromotha, kanye nokuhlaziywa kwe-SNP/InDel
Ukugcwala kokulandelana
Isichasiselo esisebenzayo sezakhi zofuzo zokubhala ikhodi
Ubudlelwano phakathi kwamasampula
Ukuhlaziywa kwe-Differential Expressed Genes (DEGs)
Ukuhlaziywa kokunothisa okusebenzayo
isichasiselo se-sRNA
Hlola intuthuko eyenziwe yizinsizakalo zokulandelana kwe-mRNA ze-BMKGene ze-Nanopore kulo magazini ofakiwe.
UGuan, CP et al. (2018) 'Izinguquko Zomhlaba Wonke Zokubhalwa Kwe-Biofilm-Forming Staphylococcus epidermidis Eziphendula Ama-Alkaloid Aphelele E-Sophorea Alopecuroides',Ijenali yasePoland yeMicrobiology, 67(2), p. 223. doi: 10.21307/PJM-2018-024.
