Metagenomic Sequencing-TGS
Metagenome ndi gulu la zinthu zopezeka m'magulu osiyanasiyana a zamoyo, monga chilengedwe ndi anthu. Lili ndi ma genomes omwe amalimidwa komanso osalimidwa. Kutsatizana kwa Metagenomic kumathandizira kusanthula kwamitundu yodabwitsayi yophatikizidwa mu zitsanzo zachilengedwe popereka zambiri kuposa mbiri ya taxonomic. Imaperekanso mawonekedwe ogwiritsira ntchito ma genomics poyang'ana majini osungidwa ndi maudindo awo pazochitika zachilengedwe. Ngakhale kuti mfuti zachikhalidwe zimayandikira ndi kutsatizana kwa Illumina zakhala zikugwiritsidwa ntchito kwambiri m'maphunziro a metagenomic, kubwera kwa Nanopore ndi PacBio kuwerengera kwanthawi yayitali kwasintha gawoli. Ukadaulo wa Nanopore ndi PacBio umakulitsa kusanthula kwa bioinformatic, makamaka msonkhano wa metagenome, kuwonetsetsa kuti misonkhano ipitirire. Malipoti akuwonetsa kuti Nanopore-based and PacBio-based metagenomics apanga bwino ma genome athunthu ndi otsekedwa a bakiteriya kuchokera ku ma microbiomes ovuta (Moss, EL, et al., Nature Biotech, 2020). Kuphatikiza mawerengedwe a Nanopore ndi mawerengedwe a Illumina kumapereka njira yowongolera zolakwika, kuchepetsa kulondola kwa Nanopore. Kuphatikizana kophatikizana kumeneku kumawonjezera mphamvu za nsanja iliyonse yotsatizana, kumapereka yankho lamphamvu kuti athe kuthana ndi malire omwe angakhalepo ndikupititsa patsogolo kulondola komanso kudalirika kwa kusanthula kwa metagenomic.
Platform: Nanopore PromethION 48, Illumia ndi PacBio Revio
Mawonekedwe a Utumiki
● Kuwerengera kwanthawi yayitali pa Nanopore P48 (laibulale ya 10Kb); PacBio Revio (laibulale ya 8Kb)
● Kukonza zolakwika: kutsatizana pa Illumina NovaSeq (PE150 library)
Ubwino wa Utumiki
●Msonkhano Wapamwamba wa Metagenome:Ndi ma contig ochepa omwe ali ndi kupitiliza kwapamwamba.
●Kulondola Kwambiri:Kuzindikiritsa mitundu ndi kuneneratu kwa jini.
●Msonkhano Wotukuka: Kuthandizira kudzipatula kwa ma genome a bakiteriya komanso kusanthula kwa metagenome.
●Comprehensive Bioinformatics Analysis:Kusanthula kwathu sikungoyang'ana pa kusiyana kwa taxonomic komanso kusiyanasiyana kwa anthu ammudzi.
●Zochitika Zazambiri: Gulu lathu limabweretsa zokumana nazo zambiri pantchito iliyonse, yokhala ndi mbiri yotseka bwino ma projekiti angapo a metagenomics m'magawo osiyanasiyana ofufuza ndikukonza zitsanzo zopitilira 200,000, gulu lathu limabweretsa zokumana nazo zambiri pantchito iliyonse.
Mafotokozedwe a Utumiki
| Sequencing nsanja | Njira Yotsatirira | Deta yovomerezeka |
| Illumina NovaSeq | Mtengo wa PE150 | 6gb pa |
| Nanopore P48 | 10 kb ku | 6/10 Gb |
| PacBio Revio | 8kb pa | 10/20 Gb |
Zitsanzo Zofunika
| Kukhazikika (ng/µL) | Ndalama zonse (µg) | Voliyumu (µL) | |
| Library ya Illumina PE150 | ≥1 | ≥0.03 | ≥20 |
| Nanopore 10 kb library | ≥40 | ≥2 | ≥20 |
| Laibulale ya PacBio 8 kb | ≥50 | 10 μg / selo | ≥20 |
Kuyenda kwa Ntchito Yantchito
Kupereka zitsanzo
Kumanga laibulale
Kutsata
Kusanthula deta
Pambuyo pogulitsa ntchito
Mulinso zowunikira izi:
● Kutsata Kuwongolera kwa Ubwino wa Data
● Metagenome Assembly ndi Gene Prediction
● Mawu a Gene
● Taxonomic Alpha Diversity Analysis
● Kusanthula kwa Ntchito Zamagulu: Biological Function, Metabolic, Antibiotic Resistance
● Kuwunika pa Zosiyanasiyana za Functional ndi Taxonomic:
Beta Diversity Analysis
Kusanthula kwamagulu
Kusanthula Kwamalumikizidwe: pakati pa zinthu zachilengedwe ndi kapangidwe ka OUT ndi kusiyanasiyana
Kugawa ma jini osafunikira:
Ndemanga za Gene: KEGG njira
Ndemanga za Gene: eggNOG
Kufotokozera kwa gene: CAZY carbohydrates
Netiweki yolumikizana ndi mitundu:
Onani kupita patsogolo komwe kumayendetsedwa ndi BMKGene's metagenomics service ndi Nanopore + Illumina ndi cholembedwa ichi:
Jin, H. et al. (2023) 'A high-quality genome compendium of the human gut microbiome of Inner Mongolians', Nature Microbiology 2023 8: 1, 8 (1), pp. 150-161. doi: 10.1038/s41564-022-01270-1.
