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Metagenomic Sequencing-Nanopore

Metagenomics chishandiso chemolecular chinoshandiswa kuongorora zvakasanganiswa genomic zvinhu zvakatorwa kubva kune zvakatipoteredza samples, izvo zvinopa ruzivo rwakadzama mukusiyana kwemarudzi uye kuwanda, chimiro chehuwandu hwevanhu, phylogenetic relationship, functional genes uye correlation network nezvakatipoteredza, etc. Nanopore sequencing platforms ichangobva kuiswa kune zvidzidzo zvemetagenomic.Kuita kwayo kwakatanhamara mukureba kwekuverenga kwakawedzera kudzika pasi metagenomic ongororo, kunyanya metagenome musangano.Kutora mabhenefiti ekuverenga-kureba, Nanopore-yakavakirwa metagenomic kudzidza inokwanisa kuwana yakawanda inoenderera gungano ichienzanisa nepfuti-pfuti metagenomics.Yakaburitswa kuti Nanopore-yakavakirwa metagenomics yakabudirira kuburitsa yakazara uye yakavharwa mabhakitiriya genomes kubva kumamicrobiomes (Moss, EL, et. al,Nature Biotech, 2020)

Platform:Nanopore PromethION P48


Service Details

Demo Results

Nyaya yeBMK

Zvakanakira Basa

ØKuungana kwepamusoro-Kuwedzera kurongeka kwekuzivikanwa kwezvipenyu uye kufanotaura kwemajini funcational

ØYakavharwa bhakitiriya genome isolation

ØYakawedzera simba uye yakavimbika application munzvimbo dzakasiyana siyana, semuenzaniso kucherechedzwa kwemapathogenic microorganisms kana maantibiotic resistance ane chekuita nemajini.

ØKuenzanisa metagenome kuongorora

Sevhisi Zvinotsanangurwa

SequencingPlatform

Library

Inokurudzirwa kuburitsa data

Inofungidzirwa kutendeuka-nguva

Illumina NovaSeq 6000

PE250

50K/100K/300K Tags

30 Mazuva

Bioinformatics inoongorora

üRaw data quality control

üMetagenome musangano

üNon-redundant gene set uye annotation

üKuongororwa kwemhando dzakasiyana-siyana

üGenetic function diversity analysis

üInter-group analysis

üKuongororwa kwemubatanidzwa kunopesana nekuedza zvinhu

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Sample Zvinodiwa uye Delivery

Sample zvinodiwa uye kuendesa

Sample Zvinodiwa:   

ForDNA zvinyorwa:

Muenzaniso Type

Mari

Concentration

Kuchena

DNA zvinyorwa

> 30 ng

> 1 ng/μl

OD260/280= 1.6-2.5

Zvezvakatipoteredza samples:

Muenzaniso mhando

Inokurudzirwa sampling maitiro

Ivhu

Sampling huwandu: approx.5 g;Chinhu chakasara chakasvava chinoda kubviswa pamusoro;Geya zvidimbu zvakakura uye upfuure ne2 mm sefa;Aliquot samples mune isingabereki EP-chubhu kana cyrotube yekuchengetera.

Feces

Sampling huwandu: approx.5 g;Unganidza uye aliquot samples mune isina utachiona EP-tube kana cryotube yekuchengetera.

Zviri mukati mematumbo

Samples dzinofanirwa kugadziriswa pasi peaseptic mamiriro.Geza matishu akaunganidzwa nePBS;Centrifuge iyo PBS uye tora iyo precipitant muEP-machubhu.

Sludge

Sampling huwandu: approx.5 g;Unganidza uye aliquot sludge sampuli mune isina utachiona EP-tube kana cryotube yekuchengetera

Mvura

Yemuenzaniso ine hushoma hushoma hwehutachiona, senge mvura yemupombi, mvura yemugodhi, nezvimwewo, Unganidza ingangoita 1 L mvura uye upfuure nepakati pe 0.22 μm sefa yekupfumisa hutachiona pa membrane.Chengetedza membrane muchubhu isina utachiona.

Ganda

Nyatsokwenya ganda nedonje swab isina utachiona kana blade yekuvhiya woiisa muchubhu isina utachiona.

Yakakurudzirwa Sample Delivery

Svitsa masampula munitrogen yemvura kwemaawa 3-4 uye chengetedza mune yemvura nitrogen kana -80 dhigirii kusvika kureba kureba.Sample shipping with dry-ice inodiwa.

Kuyerera Kwebasa Rebasa

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Muenzaniso wekutumira

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Kuvakwa kweraibhurari

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Sequencing

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Data analysis

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Mushure mekutengesa masevhisi


  • Zvakapfuura:
  • Zvinotevera:

  • 1.Heatmap: Species hupfumi kuunganidza32.Magenes anoshanda anotsanangurwa kuKEGG nzira dzemagetsi43.Species correlation network54.Circos yeCARD antibiotic resistance genes
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    Nyaya yeBMK

    Nanopore metagenomics inogonesa kukurumidza kuongororwa kwekiriniki yehutachiona hwehutachiona hwepazasi

    Rakatsikiswa:Nature Biotechnology, 2019

    Zvinokosha Zveunyanzvi
    Sequencing: Nanopore MinION
    Clinical metagenomics bioinformatics: Host DNA kupera, WIMP uye ARMA kuongorora.
    Kukurumidza kuona: maawa matanhatu
    Kunzwa kwakanyanya: 96.6%

    Mibairo yakakosha

    Muna 2006, chirwere chepasi chekufema (LR) chakakonzera kufa kwevanhu mamiriyoni matatu pasi rose.Iyo yakajairwa nzira yeLR1 pathogen yekuona kurima, iyo ine kusanzwa zvakanaka, kutendeuka-nguva-kutenderera-nguva uye kushomeka kwekutungamira mukutanga kwemishonga inorwisa mabhakitiriya.Kukurumidza uye kwakarurama kuongororwa kwehutachiona hwagara huri chido chekukurumidzira.Dr. Justin kubva kuYunivhesiti yeEast Anglia nevamwe vake vakabudirira kugadzira nzira yeNanopore-based metagenomic yekutsvaga utachiona.Zvinoenderana nekufamba kwavo, 99.99% yevaenzi DNA inogona kudzikiswa.Kuonekwa muzvirwere uye majini asingachinjiki nemishonga inorwisa mabhakitiriya anogona kupera mumaawa matanhatu.

    Reference
    Charalampous, T. , Kay, GL , Richardson, H. , Aydin, A. , & O'Grady, J. .(2019).Nanopore metagenomics inogonesa kukurumidza kuongororwa kwekiriniki yehutachiona hwehutachiona hwepazasi.Nature Biotechnology, 37(7), 1.

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