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Metagenomic Sequencing (NGS)

Metagenome (Metagenome) inoreva muunganidzwa wezvakakwana zvemageneti emusanganiswa wenharaunda yezvipenyu, senge yezvakatipoteredza metagenome, yemunhu metagenome, nezvimwewo. Ine majenomes ezvose zvinorimwa uye zvisingarimike tupukanana.Metagenomic sequencing is a molecular tool inoshandiswa kuongorora zvakasanganiswa genomic zvinhu zvakatorwa kubva kune zvakatipoteredza samples, izvo zvinopa ruzivo rwakadzama mumhando dzakasiyana-siyana uye kuwanda, chimiro chehuwandu hwevanhu, hukama hwephylogenetic, kushanda kwemajini uye correlation network ine zvakatipoteredza.

Platform:Illumina NovaSeq6000


Service Details

Demo Results

Nyaya Yokudzidza

Zvakanakira Basa

ØKuzviparadzanisa nevamwe uye kurima-yemahara kune microbial nharaunda profiling

ØKugadziriswa kwepamusoro mukuona mhando dzemhando dzakaderera mumasampuli ezvakatipoteredza

ØPfungwa ye "meta-" inobatanidza zvinhu zvose zvehupenyu pahutano hwekushanda, rudzi rwemhando uye gene level, iyo inoratidza maonero ane simba ari pedyo nechokwadi.

ØBMK inounganidza ruzivo rwakakura mumhando dzakasiyana dzemasampuli ane anopfuura zviuru gumi masampuli akagadziriswa.

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:

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.Histogram: Kugoverwa kwemarudzi

    3

    2.Magenes anoshanda anotsanangurwa kuKEGG nzira dzemagetsi

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    3.Heat mepu: Mabasa akasiyana zvichienderana nehuwandu hwemajini54.Circos yeCARD antibiotic resistance genes

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    Nyaya yeBMK

    Kuwanda kwemajini ekudzivirira kwemishonga inorwisa mabhakitiriya uye hutachiona hwehutachiona pamwe nevhu-mangrove midzi inoenderera.

    Rakatsikiswa:Chinyorwa cheHazardous Materials, 2021

    Sequencing strategy:

    Zvishandiso: Zvikamu zveDNA zvezvikamu zvina zvemangrove midzi inosanganiswa masamples: ivhu risina kusimwa, rhizosphere, episphere uye endosphere compartments.
    Platform: Illumina HiSeq 2500
    Zvinangwa: Metagenome
    16S rRNA gene V3-V4 dunhu

    Mibairo yakakosha

    Metagenomic sequencing uye metabarcoding profiling paivhu-mudzi kuenderera mberi kwemangrove saplings yakagadziriswa kuti iongorore kuparadzirwa kwemishonga inorwisa mabhakitiriya (ARGs) kubva muvhu kuita zvirimwa.Metagenomic data yakaratidza kuti 91.4% yemishonga inorwisa mabhakitiriya yaiwanzozivikanwa muzvikamu zvina zvevhu zvataurwa pamusoro apa, izvo zvakaratidza fashoni inopfuurira.16S rRNA amplicon sequencing yakagadzira 29,285 sequences, inomiririra 346 marudzi.Kusanganiswa nemhando dzemhando dzemhando neamplicon sequencing, kuparadzirwa uku kwakawanikwa kwakasununguka kubva kune midzi-inosanganiswa microbiota, zvisinei, inogona kufambiswa nenharembozha yemajini.Chidzidzo ichi chakaratidza kuyerera kweArGs neutachiona kubva muvhu kupinda muzvirimwa kuburikidza nekubatana kwevhu-midzi inoenderera.

    Reference

    Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Kuwanda kwemajini ekudzivirira kwemishonga inorwisa mabhakitiriya uye utachiona hwehutachiona padyo nevhu-mangrove midzi inoenderera.Nyaya yeHazardous Materials, 408, 124985.

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