16S/18S/ITS Amplicon Sequencing-NGS
Zinthu Zofunika pa Utumiki
● Nsanja yotsatizana: Illumina NovaSeq.
● Kukulitsa madera afupi a 16S, 18S ndi ITS, pakati pa zolinga zina zokulitsa.
● Zosankha zosinthasintha za amplicon.
● Zomwe zidachitika kale pa ntchito yokhala ndi zolinga zingapo zokulitsa.
Ubwino wa Utumiki
●Palibe kudzipatula:Kuzindikira mwachangu kapangidwe ka tizilombo toyambitsa matenda m'zitsanzo zachilengedwe.
●Kukongola Kwambiri: Mu zigawo zochepa zomwe zili mu zitsanzo zachilengedwe.
●Imagwiritsidwa Ntchito Kwambiri: Maphunziro osiyanasiyana a m'dera la tizilombo toyambitsa matenda.
●Kusanthula Kwathunthu kwa Bioinformatics: Phukusi laposachedwa la QIIME2 (chidziwitso cha kuchuluka kwa zinthu zachilengedwe) lokhala ndi kusanthula kosiyanasiyana pankhani ya database, kufotokozera, OTU/ASV.
●Ukatswiri Wochuluka: Ndi mapulojekiti 150,000 amplicon sequencing omwe amachitidwa pachaka, BMKGENE imabweretsa chidziwitso cha zaka zoposa khumi, gulu lofufuza laluso kwambiri, zomwe zili mkati, komanso chithandizo chabwino kwambiri pambuyo pa malonda.
Mafotokozedwe a Utumiki
| Laibulale | Njira Yotsatizana | Deta ikulangizidwa |
| Amplicon | Illumina PE250 | Ma tag 50/100/300K (Werengani Ma Pairs) |
Zofunikira pa Utumiki
| Kuchuluka kwa madzi (ng/µL) | Chiwerengero chonse (ng) | Voliyumu (µL) |
| ≥1 | ≥200 | ≥20 |
● Dothi/matope: 1-2g
● Kuchuluka kwa matumbo - nyama: 0.5-2g
● Zomwe zili m'matumbo - tizilombo: 0.1-0.25g
● Pamwamba pa chomera (dothi lolemera): 0.1-0.5g
● Msuzi wophika bwino matope: 0.1-0.5g
● Ndowe (nyama zazikulu): 0.5-2g
● Ndowe (mbewa): timbewu 3-5
● Madzi osambira m'mapapo: pepala losefera
● Nsalu yopukutira m'mimba: Nsalu 5-6
● Nsalu yothira pakhungu/kumaliseche/malovu/minofu yofewa ya mkamwa/nsalu yothira pakhosi/nsalu yothira pakhosi: nsalu 2-3
● Tizilombo toyambitsa matenda pamwamba: pepala losefera
● Madzi/mpweya/biofilm: pepala losefera
● Ma Endophyte: 1-2g
● Chikwangwani cha Mano: 0.5-1g
Kayendedwe ka Ntchito Yotumikira
Kutumiza zitsanzo
Kumanga laibulale
Kutsatana
Kusanthula deta
Ntchito zogulitsa pambuyo pogulitsa
Kusanthula kotsatiraku kumaphatikizapo:
- Kuwongolera khalidwe la deta yosaphika
- Kusonkhanitsa kwa OTU / Kuchotsa phokoso (ASV)
- Chidule cha OTU
- Kusanthula kwa kusiyana kwa Alpha: ma index angapo, kuphatikiza Shannon, Simpson ndi ACE.
- Kusanthula kwa mitundu yosiyanasiyana ya beta
- Kusanthula pakati pa magulu
- Kusanthula kwa mgwirizano: pakati pa zinthu zachilengedwe ndi kapangidwe ndi kusiyanasiyana kwa OUT
- Kuneneratu kwa majini ogwira ntchito a 16S
Histogram ya kugawa kwa taxonomic
mapu a kutentha kwa kuchuluka kwa taxonomic
Kusanthula kwa kusiyana kwa Alpha: rarefaction curve
Kusanthula kwa mitundu yosiyanasiyana ya beta: NMDS
Kusanthula kwa magulu osiyanasiyana: Kupezeka kwa biomarker ya LEFSE
Fufuzani kupita patsogolo komwe kwathandizidwa ndi ntchito za BMKGene zotsatizana ndi Illumina kudzera mu zofalitsa zosiyanasiyana.
Dong, C. ndi ena. (2022) 'Kusonkhana, Core Microbiota, ndi Ntchito ya Rhizosphere Soil ndi Bark Microbiota mu Eucommia ulmoides', Frontiers in Microbiology, 13. doi: 10.3389/FMICB.2022.855317/FULL.
Li, Y. et al. (2023) 'Kusamutsira mabakiteriya opangidwa ndi mabakiteriya kuti athetse matenda a 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) 'Tizilombo toyambitsa matenda ta fumbi la mpweya lomwe linasonkhanitsidwa panthawi yoyesera yoteteza moyo wa bioregenerative yochokera pansi pa nthaka "Lunar Palace 365"', Environmental Microbiomes, 17(1), pp. 1–20. doi: 10.1186/S40793-022-00399-0/FIGURES/8.
Yin, S. et al. (2022) 'Kuchuluka kwa majini ogwira ntchito omwe amadalira chakudya chokhudzana ndi kusintha kwa nayitrogeni komwe kumawongolera kutayika kwa nayitrogeni mu kompositi', Bioresource Technology, 361, p. 127678. doi: 10.1016/J.BIORTECH.2022.127678.
