16S/18S/ITS Amplicon Sequencing-PacBio
Izici Zesevisi
● Ipulatifomu yokulandelana: I-PacBio Revio
● Imodi yokulandelana: I-CCS (i-HiFi ifundeka)
● Ukwandiswa kwesifunda esiqondiwe kulandelwe ukuxhumanisa ama-amplicons ngasikhathi sinye ngaphambi kokulungiswa komtapo wezincwadi we-HiFi SMRT bell
Izinzuzo Zesevisi
●Isixazululo esiphezulu se-taxonomic: Tukulandelana kwe-han short-amplicon,okwenza amazinga aphezulu okuhlukaniswa kwe-OTU ezingeni lezinhlobo zezilwane aphakame.
●Ukushaya ucingo oluyisisekelo olunembile kakhulu: Ukulandelana kwemodi ye-PacBio CCS (i-HiFi ifundeka).
●Akukho ukuhlukaniswa: Ukuhlonza ngokushesha ukwakheka kwamagciwane kumasampula emvelo.
●Kusebenza kabanzi: Izifundo zomphakathi ezahlukahlukene zamagciwane.
●Ukuhlaziywa okuphelele kwe-bioinformatic: Iphakheji yakamuva ye-QIIME2 (ukuqonda okubanzi ngemvelo yamagciwane) enokuhlaziywa okuhlukahlukene ngokwesizindalwazi, isichasiselo, i-OTU/ASV.
●Ubuchwepheshe Obubanzi: Njengoba kunezinkulungwane zamaphrojekthi okulandelana kwe-amplicon aqhutshwa minyaka yonke, i-BMKGENE iletha ulwazi olungaphezu kweminyaka eyishumi, ithimba lokuhlaziya elinekhono eliphezulu, okuqukethwe okuphelele, kanye nokusekelwa okuhle kakhulu ngemuva kokuthengisa.
Imininingwane Yesevisi
| Umtapo Wolwazi | Isu Lokulandelana | Kunconywa idatha |
| I-Amplicon | I-PacBio Revio | Amathegi angu-10/30/50 K (CCS) |
Izidingo Zesampula
| Ukugxila (ng/µL) | Inani eliphelele (µg) | Ivolumu (µL) |
| ≥5 | ≥0.3 | ≥20 |
Ukulethwa Kwesampula Okunconyiwe
Faka amasampula ku-nitrogen ewuketshezi amahora angu-3-4 bese uwagcina ku-nitrogen ewuketshezi noma ku--80 degrees kuze kube yilapho ebhukwa isikhathi eside. Kudingeka ukuthunyelwa kwesampula ngeqhwa elomile.
Ukugeleza Komsebenzi Wesevisi
Ukulethwa kwesampula
Ukwakhiwa komtapo wolwazi
Ukulandelana
Ukuhlaziywa kwedatha
Izinsizakalo zangemuva kokuthengisa
Kufaka phakathi ukuhlaziywa okulandelayo:
●Ukulawulwa kwekhwalithi yedatha eluhlaza
●Ukuhlanganiswa kwe-OTU/Ukususa umsindo (ASV)
●Isichasiselo se-OTU
●Ukuhlaziywa kokwehlukahlukana kwe-Alpha: ama-index amaningi, okuhlanganisa uShannon, uSimpson kanye ne-ACE.
●Ukuhlaziywa kokwehlukahlukana kwe-Beta
●Ukuhlaziywa kwamaqembu
●Ukuhlaziywa kobudlelwano: phakathi kwezici zemvelo kanye nokwakheka kanye nokuhlukahluka kwe-OUT
●Ukubikezela izakhi zofuzo ezisebenzayo ezingu-16S
I-Histogram yokusatshalaliswa kwe-taxonomic
Umuthi we-phylogenetic wokusatshalaliswa komphakathi
Ukuhlaziywa kokwehlukahlukana kwe-Alpha: i-ACE
inkomba
Ukuhlaziywa kokwehlukahlukana kwe-Beta: i-PCoA
Ukuhlaziywa kwamaqembu ahlukene: i-ANOVA
Hlola intuthuko eyenziwe yizinsizakalo zokulandelana kwe-amplicon ze-BMKGene nge-PacBio ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
UGao, X. noWang, H. (2023) 'Ukuhlaziywa Okuqhathanisayo Kwamaphrofayili Nemisebenzi Yamagciwane eRumen Ngesikhathi Sokuzivumelanisa Nezimo Ezihlukene (Okuluhlaza Kabusha vs. Okunotshani) Ezimvwini Ze-Alpine Merino Ezinezigaba Ezimbili Zokukhula Ehlathini Lase-Alpine', Ukuvutshelwa, 9(1), ikhasi 16. doi: 10.3390/FERMENTATION9010016/S1.
ULi, S. et al. (2023) 'Ukuthwebula izinto ezimnyama ezincane enhlabathini yasehlane kusetshenziswa i-metagenomics esekelwe ku-culturomics kanye nokuhlaziywa kwesinqumo esiphezulu', i-npj Biofilms and Microbiomes 2023 9:1, 9(1), amakhasi 1–14. doi: 10.1038/s41522-023-00439-8.
Mu, L. et al. (2022) 'Imiphumela kasawoti we-fatty acid ezicini zokubilisa, ukuhlukahluka kwamagciwane kanye nokuzinza kwe-aerobic ye-silage exubile elungiselelwe ne-alfalfa, utshani belayisi kanye ne-wheat bran', Ijenali Yesayensi Yezokudla Nezolimo, 102(4), amakhasi 1475–1487. doi: 10.1002/JSFA.11482.
UYang, J. et al. (2023) 'Ukuxhumana Phakathi Kwe-Oxidative Stress Biomarkers kanye ne-Gut Microbiota Emiphumeleni Ye-Antioxidant Yezingcezu Ezivela ku-Sonchus brachyotus DC. ku-Oxazolone-Induced Intestinal Oxidative Stress ku-Adult Zebrafish', Ama-Antioxidants 2023, uMqulu 12, Ikhasi 192, 12(1), ikhasi 192. doi: 10.3390/ANTIOX12010192.
