16S/18S/ITS Amplicon Sequencing-NGS
Iimpawu zeNkonzo
● Iqonga lokulandelelanisa: Illumina NovaSeq.
● Ukwandiswa kwemimandla emifutshane ye-16S, 18S kunye ne-ITS, phakathi kwezinye iinjongo zokwandiswa.
● Ukhetho oluguquguqukayo lwe-amplicon.
● Amava eprojekthi yangaphambili eneenjongo ezininzi zokwandisa.
Iingenelo zeNkonzo
●Akukho kuhlukaniswa:Ukuchongwa ngokukhawuleza kokwakheka kweentsholongwane kwiisampuli zokusingqongileyo.
●Isisombululo Esiphezulu: Kwizinto ezincinci kakhulu kwiisampuli zokusingqongileyo.
●Isebenza kakhulu: Izifundo ezahlukeneyo zoluntu ngeentsholongwane.
●Uhlalutyo olupheleleyo lwe-Bioinformatics: Iphakheji yamva nje ye-QIIME2 (ulwazi oluchanekileyo malunga ne-microbial ecology) enohlalutyo olwahlukeneyo ngokwedathabheyisi, isichazi-magama, i-OTU/ASV.
●Ubuchule Obubanzi: Ngeeprojekthi zokulandelelana kwe-amplicon ezingama-150 amawaka ezenziwa minyaka le, i-BMKGENE izisa amava angaphezu kweshumi leminyaka, iqela lohlalutyo elinobuchule obuphezulu, umxholo opheleleyo, kunye nenkxaso ebalaseleyo emva kokuthengisa.
Iinkcukacha zeNkonzo
| Ithala leeNcwadi | Icebo Lokulandelelanisa | Idatha iyacetyiswa |
| I-Amplicon | I-Illumina PE250 | Iithegi ezingama-50/100/300K (Funda iiPairs) |
Iimfuno zeNkonzo
| Uxinzelelo (ng/µL) | Isixa-mali sisonke (ng) | Umthamo (µL) |
| ≥1 | ≥200 | ≥20 |
● Umhlaba/udaka: 1-2g
● Umthamo wamathumbu - isilwanyana: 0.5-2g
● Umxholo wamathumbu - isinambuzane: 0.1-0.25g
● Umphezulu wesityalo (inkunkuma etyebileyo): 0.1-0.5g
● Umhluzi wokubilisa otyebileyo intlenga): 0.1-0.5g
● Indle (izilwanyana ezinkulu): 0.5-2g
● Indle (igundane): 3-5 iinkozo
● Ulwelo lokuhlamba i-alveolar yemiphunga: iphepha lesihluzi
● I-swab yesisu: ii-swab ezi-5-6
● I-swab yolusu/yesini/amathe/izicubu ezithambileyo zomlomo/i-pharyngeal swab/i-rectal swab: ii-swab ezi-2-3
● Iintsholongwane ezingaphezulu: iphepha lokucoca
● Umzimba wamanzi/umoya/i-biofilm: iphepha lokucoca
● Ii-Endophytes: 1-2g
● Iplaque yamazinyo: 0.5-1g
Ukuhamba komsebenzi wenkonzo
Ukuhanjiswa kwesampulu
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Iinkonzo zasemva kokuthengisa
Kubandakanya uhlalutyo olulandelayo:
- Ulawulo lomgangatho wedatha eluhlaza
- Ukudibanisa i-OTU/Ukususa ingxolo (ASV)
- Isichazi se-OTU
- Uhlalutyo lokwahlukahluka kweAlpha: ii-indexes ezininzi, kuquka uShannon, uSimpson kunye ne-ACE.
- Uhlalutyo lokwahlukahluka kwe-Beta
- Uhlalutyo phakathi kwamaqela
- Uhlalutyo lolwalamano: phakathi kwezinto ezingqongileyo kunye nokwakheka kunye nokwahlukahluka kwe-OUT
- Uqikelelo lwe-gene olusebenzayo lwe-16S
I-Histogram yokusasazwa kwe-taxonomic
imephu yobushushu obuqukanisa ubuninzi be-taxonomic
Uhlalutyo lokwahlukahluka kweAlpha: i-rarefaction curve
Uhlalutyo lokwahlukahluka kwe-beta: i-NMDS
Uhlalutyo lwamaqela ahlukeneyo: Ukufunyanwa kwe-LEFSE biomarker
Hlola inkqubela phambili eququzelelwe ziinkonzo ze-BMKGene zokulandelelanisa i-amplicon kunye ne-Illumina ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.
UDong, C. et al. (2022) 'Ukuhlanganiswa, iCore Microbiota, kunye nomsebenzi weRhizosphere Soil kunye neBark Microbiota kwi-Eucommia ulmoides', iFrontiers in Microbiology, 13. doi: 10.3389/FMICB.2022.855317/FULL.
ULi, Y. et al. (2023) 'Ukufakelwa kwe-synthetic bacterial consortia yonyango lwe-Gardnerella vaginalis-induced bacterial vaginosis kwiimpuku', iMicrobiome, 11(1), iphe. 1–14. doi: 10.1186/s40168-023-01497-y
UYang, J., uFu, uY. kunye noLiu, H. (2022) 'Ii-Microbiome zothuli lomoya eziqokelelwe ngexesha lovavanyo lokuxhasa ubomi oluvaliweyo olusekelwe emhlabeni "iLunar Palace 365"', Environmental Microbiomes, 17(1), iphe. 1–20. doi: 10.1186/S40793-022-00399-0/FIGURES/8.
Yin, S. et al. (2022) 'Ubuninzi be-genes ezisebenzayo ezixhomekeke kwi-feedstock ezinxulumene nokuguqulwa kwe-nitrogen okulawulwayo ukulahleka kwe-nitrogen ekubumbeni', Bioresource Technology, 361, p. 127678. doi: 10.1016/J.BIORTECH.2022.127678.
