16S/18S/ITS Ukulandelelana kwe-Amplicon-NGS
Izici Zesevisi
● Ipulatifomu yokulandelana: Illumina NovaSeq.
● Ukwandiswa kwezindawo ezimfushane ze-16S, 18S kanye ne-ITS, phakathi kwezinye izinhloso zokukhulisa.
● Ukukhetha okuguquguqukayo kwe-amplicon.
● Ulwazi lwangaphambilini lwephrojekthi olunezinhloso eziningi zokukhulisa.
Izinzuzo Zesevisi
●Akunabo abantu abahlukanisayo:Ukuhlonza ngokushesha ukwakheka kwamagciwane kumasampula emvelo.
●Isixazululo Esiphezulu: Ezingxenyeni ezitholakala kalula kumasampula emvelo.
●Kusebenza Kakhulu: 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: Ngamaphrojekthi okulandelana kwe-amplicon ayi-150,000 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-Illumina PE250 | Amathegi angu-50/100/300K (Funda Amabhangqa) |
Izidingo Zesevisi
| Ukugxila (ng/µL) | Inani eliphelele (ng) | Ivolumu (µL) |
| ≥1 | ≥200 | ≥20 |
● Inhlabathi/udaka: 1-2g
● Okuqukethwe kwamathumbu - isilwane: 0.5-2g
● Okuqukethwe emathunjini - isinambuzane: 0.1-0.25g
● Ubuso besitshalo (inhlabathi ecebile): 0.1-0.5g
● Umhluzi wokubilisa ocebile nge-sediment): 0.1-0.5g
● Indle (izilwane ezinkulu): 0.5-2g
● Indle (igundane): izinhlamvu ezingu-3-5
● Uketshezi lokuhlanza i-alveolar yamaphaphu: iphepha lokuhlunga
● I-swab yesitho sangasese sowesifazane: ama-swab angu-5-6
● I-swab yesikhumba/yesitho sangasese/amathe/izicubu ezithambile zomlomo/i-swab ye-pharyngeal/i-rectal swab: ama-swab angu-2-3
● Izinto eziphilayo ezingaphezulu: iphepha lokuhlunga
● Umzimba wamanzi/umoya/i-biofilm: iphepha lokuhlunga
● Ama-Endophyte: 1-2g
● Ibhodi Lamazinyo: 0.5-1g
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: izinkomba eziningi, okuhlanganisa uShannon, uSimpson kanye ne-ACE.
- Ukuhlaziywa kokwehlukahluka kwe-Beta
- Ukuhlaziywa kwamaqembu
- Ukuhlaziywa kobudlelwano: phakathi kwezici zemvelo kanye nokwakheka kanye nokuhlukahluka kwe-OUT
- Ukubikezela kwezakhi zofuzo ezisebenzayo ze-16S
I-Histogram yokusatshalaliswa kwe-taxonomic
imephu yokushisa ehlanganisa ukugcwala kwe-taxonomic
Ukuhlaziywa kokwehlukahlukana kwe-Alpha: ijika le-rarefaction
ukuhlaziywa kokwehlukahluka kwe-beta: i-NMDS
Ukuhlaziywa kwamaqembu ahlukene: Ukutholwa kwe-biomarker ye-LEFSE
Hlola intuthuko eyenziwe yizinsizakalo zokulandelana kwe-amplicon ze-BMKGene nge-Illumina ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
UDong, C. nabanye. (2022) 'Ukuhlanganiswa, i-Core Microbiota, kanye Nomsebenzi we-Rhizosphere Soil kanye ne-Bark Microbiota ku-Eucommia ulmoides', Imingcele ku-Microbiology, 13. doi: 10.3389/FMICB.2022.855317/FULL.
ULi, Y. nabanye. (2023) 'Ukufakelwa kwe-synthetic bacterial consortia yokwelapha i-Gardnerella vaginalis-induced bacterial vaginosis emagundwini', i-Microbiome, 11(1), amakhasi 1–14. doi: 10.1186/s40168-023-01497-y
UYang, J., uFu, Y. kanye noLiu, H. (2022) 'Ama-microbiome othuli lomoya aqoqwe ngesikhathi sokuhlolwa kokusekela impilo okusekelwe emhlabathini okuvaliwe "iLunar Palace 365"', Environmental Microbiomes, 17(1), amakhasi 1–20. doi: 10.1186/S40793-022-00399-0/FIGURES/8.
Yin, S. et al. (2022) 'Ubuningi bezakhi zofuzo ezisebenzayo ezincike ekudleni okuhlobene nokuguqulwa kwe-nitrogen okulawulwa ukulahleka kwe-nitrogen ekuhlanganiseni', Bioresource Technology, 361, k. 127678. doi: 10.1016/J.BIORTECH.2022.127678.
