16S/18S/ITS Amplicon Sequencing-NGS
Izici Zesevisi
● Inkundla yokulandelanisa: Illumina NovaSeq.
● Ukukhulisa izifunda ezimfushane ze-16S, 18S ne-ITS, phakathi kokunye okuhlosiwe kokukhulisa.
● Ukukhetha okuguquguqukayo kwe-amplicon.
● Ulwazi lwephrojekthi yangaphambilini enezinjongo eziningi zokukhulisa.
Izinzuzo Zesevisi
●Ukungahlukaniswa muntu:Ukuhlonzwa okusheshayo kokwakheka kwe-microbial kumasampuli emvelo.
●Ukulungiswa Okuphezulu: Ezingxenyeni ezinenala ephansi kumasampuli emvelo.
●Kusebenza Ngokubanzi: Izifundo ezihlukene ze-microbial community.
●Ukuhlaziywa Kwe-Bioinformamatic Comprehensive: Iphakheji yakamuva ye-QIIME2 (imininingwane yobuningi ku-microbial ecology) enokuhlaziya okuhlukahlukene ngokuya kusizindalwazi, isichasiselo, i-OTU/ASV.
●Ubungcweti Olubanzi: Ngamaphrojekthi wokulandelanisa ama-amplicon ayizinkulungwane eziyi-150 aqhutshwa minyaka yonke, i-BMKGENE iletha isipiliyoni seminyaka engaphezu kweshumi, ithimba lokuhlaziya elinekhono eliphezulu, okuqukethwe okuphelele, nokusekelwa okuhle kakhulu kwangemuva kokuthengisa.
Imininingwane Yesevisi
| Umtapowolwazi | Isu Lokulandelanisa | Idatha iyanconywa |
| I-Amplicon | I-Ilumina PE250 | 50/100/300K omaka (Funda ngababili) |
Izidingo Zesevisi
| Ukugxila (ng/µL) | Inani eliphelele (ng) | Ivolumu (µL) |
| ≥1 | ≥200 | ≥20 |
● Umhlabathi/udaka: 1-2g
● Okuqukethwe kwamathumbu-isilwane: 0.5-2g
● Okuqukethwe kwamathumbu-insect: 0.1-0.25g
● Indawo yezitshalo (i-sediment enothile): 0.1-0.5g
● Umhluzi ovutshelwayo ucebisile inzika): 0.1-0.5g
● Indle (izilwane ezinkulu): 0.5-2g
● Indle (igundane): 3-5grains
● I-pulmonary alveolar lavage fluid: iphepha lokuhlunga
● I-swab yesitho sangasese sowesifazane: 5-6 swabs
● Isikhumba/i-swab yezitho zangasese/amathe/izicubu ezithambile zomlomo/i-pharyngeal swab/i-rectal swab: 2-3 swabs
● Ama-microorganisms angaphezulu: iphepha lokuhlunga
● I-Waterbody/air/biofilm: isihlungi sephepha
● Ama-Endophyte: 1-2g
● Uqweqwe Lwamazinyo: 0.5-1g
Ukugeleza Komsebenzi Wesevisi
Ukulethwa kwesampula
Ukwakhiwa komtapo wolwazi
Ukulandelanisa
Ukuhlaziywa kwedatha
Amasevisi angemuva kokuthengisa
Kufaka ukuhlaziya okulandelayo:
- Ukulawulwa kwekhwalithi yedatha eluhlaza
- Ukuhlanganisa kwe-OTU /De-noise(ASV)
- OTU isichasiselo
- Ukuhlaziywa kokuhlukahluka kwe-Alpha: izinkomba eziningi, okuhlanganisa i-Shannon, i-Simpson ne-ACE.
- Ukuhlaziywa kokuhlukahluka kwe-Beta
- Ukuhlaziya kwamaqembu
- Ukuhlaziywa kokuhlobana: phakathi kwezici zemvelo kanye nokwakheka kwe-OUT nokuhlukahluka
- 16S isibikezelo sofuzo esisebenzayo
I-Histogram yokusabalalisa kwe-taxonomic
imephu yokushisa ehlanganisayo ye-taxonomic
Ukuhlaziywa kokuhlukahluka kwe-Alpha: ijika le-rarefaction
Ukuhlaziywa kokuhlukahluka kwe-beta: NMDS
Ukuhlaziywa kwe-Intergroup: ukutholwa kwe-biomarker ye-LEFSE
Hlola intuthuko egqugquzelwa izinsiza zokulandelana kweamplicon ye-BMKGene ne-Illumina ngeqoqo elikhethiwe lokushicilelwe.
Dong, C. et al. (2022) 'Assembly, Core Microbiota, and Function of the Rhizosphere Soil and Bark Microbiota in Eucommia ulmoides', Frontiers in Microbiology, 13. doi: 10.3389/FMICB.2022.855317/FULL.
Li, Y. et al. (2023) 'I-Synthetic bacterial consortia transplantation yokwelashwa kwe-Gardnerella vaginalis-induced bacterial vaginosis kumagundane', i-Microbiome, i-11 (1), iphe. 1-14. doi: 10.1186/s40168-023-01497-y
U-Yang, J., Fu, Y. kanye no-Liu, H. (2022) 'Ama-Microbiomes othuli lomoya aqoqwe ngesikhathi sokuhlolwa kokusekelwa okusekelwe phansi kwe-biogenerative yokuphila “Lunar Palace 365”', Environmental Microbiomes, 17(1), pp. 1–20. doi: 10.1186/S40793-022-00399-0/AMAFIGURES/8.
Yin, S. et al. (2022) 'Ubuningi obuncike ku-Feedstock kwezakhi zofuzo ezisebenzayo ezihlobene nokuguqulwa kwe-nitrogen kulawulwa ukulahleka kwe-nitrogen ekwenzeni umquba', I-Bioresource Technology, 361, p. 127678. doi: 10.1016/J.BIORTECH.2022.127678.
