16S/18S/ITS Amplicon Sequencing-NGS
Mawonekedwe a Utumiki
● Sequencing platform: Illumina NovaSeq.
● Kukulitsa zigawo zazifupi za 16S, 18S ndi ITS, pakati pa zolinga zina zokulitsa.
● Zosankha zosinthika za amplicon.
● Zokumana nazo m'mbuyomo za polojekiti yokhala ndi zolinga zingapo zokulitsa.
Ubwino wa Utumiki
●Zopanda kudzipatula:Chizindikiritso chofulumira cha kapangidwe ka tizilombo tating'onoting'ono mu zitsanzo zachilengedwe.
●Kukhazikika Kwambiri: Mu zigawo zotsika kwambiri mu zitsanzo zachilengedwe.
●Yogwiritsidwa Ntchito Kwambiri: Maphunziro osiyanasiyana amagulu a tizilombo.
●Comprehensive Bioinformatic Analysis: Phukusi laposachedwa la QIIME2 (kachulukidwe kazinthu zachilengedwe) yokhala ndi kusanthula kosiyanasiyana malinga ndi nkhokwe, ndemanga, OTU/ASV.
●Katswiri Wazambiri: Ndi ntchito 150 zikwi amplicon sequencing zomwe zimachitika chaka ndi chaka, BMKGENE imabweretsa zaka khumi zachidziwitso, gulu lofufuza laluso kwambiri, zokhutira, ndi chithandizo chabwino kwambiri cha pambuyo pa malonda.
Mafotokozedwe a Utumiki
| Library | Njira Yotsatirira | Deta yovomerezeka |
| Amplicon | Illumina PE250 | 50/100/300K ma tag (Werengani awiriawiri) |
Zofunikira pa Utumiki
| Kukhazikika (ng/µL) | Ndalama zonse (ng) | Voliyumu (µL) |
| ≥1 | ≥200 | ≥20 |
● Dothi / matope: 1-2g
● Zinyama za m'mimba: 0.5-2g
● Zomwe zili m'matumbo-tizilombo: 0.1-0.25g
● Pamwamba pa chomera (chidothi chowonjezera): 0.1-0.5g
● Msuzi wowotchera wowonjezera matope): 0.1-0.5g
● Nyasi (zinyama zazikulu): 0,5-2g
● Nyasi (mbewa): 3-5mbewu
● Pulmonary alveolar lavage fluid: pepala losefera
● Kupukuta maliseche: 5-6 swabs
● Khungu/kumaliseche/kumaliseche/malovu/mnofu wofewa m'kamwa/pharyngeal swab/nsomba: 2-3 swabs
● Tizilombo tating'onoting'ono: pepala losefera
● Waterbody/air/biofilm: pepala losefera
● Endophytes: 1-2g
● Mpanda Wamano: 0.5-1g
Kuyenda kwa Ntchito Yantchito
Kupereka zitsanzo
Kumanga laibulale
Kutsata
Kusanthula deta
Pambuyo pogulitsa ntchito
Mulinso zowunikira izi:
- Kuwongolera khalidwe la data yaiwisi
- Magulu a OTU / De-noise (ASV)
- Chidziwitso cha OTU
- Kusanthula kwamitundu ya Alpha: ma index angapo, kuphatikiza Shannon, Simpson ndi ACE.
- Kusanthula kwamitundu yosiyanasiyana ya Beta
- Kusanthula kwamagulu
- Kusanthula kwa mgwirizano: pakati pa zinthu zachilengedwe ndi kapangidwe ka OUT ndi kusiyanasiyana
- 16S magwiridwe antchito a jini
Histogram ya kugawa taxonomic
taxonomic abundance clustering heat map
Kusanthula kwamitundu ya Alpha: rarefaction curve
Kusanthula kwamitundu yosiyanasiyana ya beta: NMDS
Kusanthula kwamagulu: LEFSE biomarker anapeza
Onani kupita patsogolo komwe kumayendetsedwa ndi BMKGene's amplicon sequencing services ndi Illumina kudzera mgulu lazofalitsa.
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) 'Synthetic bacterial consortia transplantation pofuna kuchiza Gardnerella vaginalis-induced bacterial vaginosis mu mbewa', Microbiome, 11 (1), pp. 1-14. doi: 10.1186/s40168-023-01497-y
Yang, J., Fu, Y. ndi Liu, H. (2022) 'Microbiomes of air fumbi anasonkhanitsidwa panthawi yapansi yotsekedwa yotseka biogenerative moyo kuyesa "Lunar Palace 365"', Environmental Microbiomes, 17(1), pp. 1–20. doi: 10.1186/S40793-022-00399-0/ZINTHU/8.
Yin, S. et al. (2022) 'Kuchuluka kodalira kwa feedstock kwa majini ogwira ntchito okhudzana ndi kusintha kwa nayitrogeni kumayang'anira kutaya kwa nayitrogeni mu kompositi', Bioresource Technology, 361, p. 127678. doi: 10.1016/J.BIORTECH.2022.127678.
