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Zogulitsa

Metagenomic Sequencing (NGS)

Metagenome imatanthawuza kusonkhanitsa kwa chibadwa chamagulu osakanikirana a zamoyo monga chilengedwe, metagenome yaumunthu, ndi zina zotero. Lili ndi ma genome a tizilombo tomwe timalima komanso tosalimidwa.Metagenomic sequencing ndi chida cha mamolekyu chomwe chimagwiritsidwa ntchito posanthula zinthu zosakanikirana zamtundu wotengedwa kuchokera ku zitsanzo zachilengedwe, zomwe zimapereka chidziwitso chambiri pamitundu yosiyanasiyana ndi kuchuluka kwake, kuchuluka kwa anthu, ubale wa phylogenetic, majini ogwira ntchito komanso maukonde olumikizana ndi chilengedwe.

nsanja:Illumina NovaSeq6000


Tsatanetsatane wa Utumiki

Zotsatira za Demo

Nkhani Yophunzira

Ubwino wa Utumiki

ØKudzipatula komanso kulima kopanda mbiri yamagulu ang'onoang'ono

ØKusamvana kwakukulu pakuzindikira mitundu yocheperako mu zitsanzo zachilengedwe

ØLingaliro la "meta-" limagwirizanitsa zinthu zonse zamoyo pamlingo wogwirira ntchito, mlingo wa mitundu ndi jini, zomwe zimasonyeza malingaliro amphamvu omwe ali pafupi ndi zenizeni.

ØBMK imadziunjikira zambiri mumitundu yosiyanasiyana yokhala ndi zitsanzo zopitilira 10,000 zokonzedwa.

Mafotokozedwe a Utumiki

Kutsatansanja

Library

Zokolola zovomerezeka

Nthawi yoyerekeza yozungulira

Illumina NovaSeq 6000

Chithunzi cha PE250

50K/100K/300K Tags

Masiku 30

Bioinformatics kusanthula

üKuwongolera khalidwe la data yaiwisi

üMsonkhano wa Metagenome

üMa jini osagwirizana ndi ma jini ndi ndemanga

üKusanthula kwamitundu yosiyanasiyana

üKusanthula kwamitundu yosiyanasiyana ya ma genetic

üKusanthula kwamagulu

üKusanthula kwamagulu motsutsana ndi zinthu zoyeserera

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Zitsanzo Zofunika ndi Kutumiza

Zitsanzo Zofunika:

ZaZithunzi za DNA:

Mtundu Wachitsanzo

Ndalama

Kukhazikika

Chiyero

Zithunzi za DNA

> 30 ng

> 1 ng/μl

OD260/280= 1.6-2.5

Za zitsanzo zachilengedwe:

Mtundu wachitsanzo

Njira yoyeserera yolangizidwa

Nthaka

Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Chotsalira chofota chiyenera kuchotsedwa pamwamba;Pewani zidutswa zazikulu ndikudutsa fyuluta ya 2 mm;Zitsanzo za Aliquot mu chubu chosabala cha EP kapena cyrotube kuti zisungidwe.

Ndowe

Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Sonkhanitsani ndi aliquot zitsanzo mu EP-chubu wosabala kapena cryotube kuti musungidwe.

Zomwe zili m'matumbo

Zitsanzo ziyenera kukonzedwa pansi pa chikhalidwe cha aseptic.Sambani minofu yosonkhanitsidwa ndi PBS;Centrifuge the PBS ndi kusonkhanitsa precipitant mu EP-machubu.

Sludge

Kuchuluka kwa zitsanzo: pafupifupi.5 g pa;Sungani ndi aliquot sludge zitsanzo mu wosabala EP-chubu kapena cryotube kusungitsa

Madzi

Kwa zitsanzo zokhala ndi tizilombo tating'onoting'ono, monga madzi apampopi, madzi apachitsime, ndi zina zotero, Sonkhanitsani madzi osachepera 1 L ndikudutsa 0.22 μm fyuluta kuti mulemeretse tizilombo toyambitsa matenda pa nembanemba.Sungani nembanemba mu chubu chosabala.

Khungu

Pala pamwamba pa khungu ndi thonje wosabala kapena tsamba la opaleshoni ndikuyika mu chubu wosabala.

Kutumiza Kwachitsanzo Kovomerezeka

Ikani zitsanzo mu nayitrogeni wamadzimadzi kwa maola 3-4 ndikusunga mu nayitrogeni wamadzimadzi kapena -80 digiri kuti musungidwe nthawi yayitali.Kutumiza kwachitsanzo kokhala ndi ayezi wowuma ndikofunikira.

Kuyenda kwa Ntchito Yantchito

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Kutumiza kwachitsanzo

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Kumanga laibulale

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Kutsata

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Kusanthula deta

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Pambuyo pogulitsa ntchito


  • Zam'mbuyo:
  • Ena:

  • 1.Histogram: Kugawa kwamitundu

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    2.Majini ogwira ntchito ofotokozera njira za metabolic za KEGG

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    Mapu a 3.Kutentha: Ntchito zosiyana zotengera kuchuluka kwa majini54.Circos of CARD antibiotic resistance genenes

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    BMK Case

    Kuchuluka kwa majini olimbana ndi maantibayotiki ndi tizilombo toyambitsa matenda m'mphepete mwa mizu ya mangrove

    Lofalitsidwa:Journal of Hazardous Equipment, 2021

    Njira yotsatirira:

    Zipangizo: Zotulutsa za DNA za zidutswa zinayi za mizu ya mangrove yokhudzana ndi zitsanzo: nthaka yosabzalidwa, rhizosphere, episphere ndi endosphere compartments
    Pulatifomu: Illumina HiSeq 2500
    Zolinga: Metagenome
    16S rRNA jini V3-V4 dera

    Zotsatira zazikulu

    Tsatanetsatane wa metagenomic ndi mbiri ya metabarcoding pamizu yopitilira muzu wa mitengo ya mangrove adakonzedwa kuti aphunzire kufalitsa ma antibiotic resistance genes (ARGs) kuchokera kunthaka kupita ku zomera.Deta ya Metagenomic inavumbulutsa kuti 91.4% ya majini olimbana ndi maantibayotiki amadziwika bwino m'zigawo zonse zinayi za dothi zomwe zatchulidwa pamwambapa, zomwe zimasonyeza kachitidwe kosalekeza.16S rRNA amplicon sequencing adapanga 29,285 motsatira, kuyimira mitundu 346.Kuphatikizana ndi mbiri ya zamoyo pogwiritsa ntchito ma amplicon sequencing, kufalitsa kumeneku kunapezeka kuti sikunachokere ku microbiota yokhudzana ndi mizu, komabe, ikhoza kuthandizidwa ndi mafoni amtundu.Kafukufukuyu adazindikira kutuluka kwa ma ARGs ndi tizilombo toyambitsa matenda kuchokera m'nthaka kulowa muzomera kudzera munjira yolumikizana ndi mizu ya nthaka.

    Buku

    Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Kuchuluka kwa majini olimbana ndi maantibayotiki ndi tizilombo toyambitsa matenda m'nthaka-mizu ya mangrove mosalekeza.Journal of Hazardous Materials, 408, 124985.

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