BMKCloud Log in
条形 fu'a-03

Oloa

Metagenomic Sequencing -NGS

Metagenome e fa'atatau i le aofa'i o mea fa'atupu o se fa'alapotopotoga fefiloi o meaola, e pei o le si'osi'omaga metagenome, tagata metagenome, ma isi. O lo'o iai genomes o meaola ninii e mafai ona fa'ato'aina ma mea e le mafai ona fa'ato'aina.Metagenomic sequencing ose meafaigaluega mole faʻaaogaina e suʻesuʻe ai mea faʻafefiloi genomic na maua mai faʻataʻitaʻiga o le siosiomaga, lea e maua ai faʻamatalaga auiliili i ituaiga eseese ma le tele, fausaga o le faitau aofaʻi, sootaga phylogenetic, genes galue ma fesoʻotaʻiga fesoʻotaʻiga ma mea tau le siosiomaga.

Fa'avae:Illumina NovaSeq Platform


Au'aunaga Fa'amatalaga

I'uga Fa'aaliga

Tulaga Suesuega

Tulaga Auaunaga

● Tu'u'ese ma fa'ato'aga e leai se totogi mo fa'amatalaga fa'alaua'itele a microbial

● Maualuluga maualuga i le suʻeina o ituaiga e maualalo le tele i faʻataʻitaʻiga o le siosiomaga

● O le manatu o le "meta-" o loʻo tuʻufaʻatasia uma uiga faʻaola i le tulaga galue, tulaga o ituaiga ma le gene, lea e atagia mai ai se vaaiga malosi e latalata i le mea moni.

● BMK faʻaputuina le poto masani i ituaiga faʻataʻitaʻiga eseese ma sili atu i le 10,000 faʻataʻitaʻiga faʻatautaia.

Fa'amatalaga Auaunaga

 Fa'avae

Fa'asologa

Fa'amatalaga fautuaina

Taimi liliu

Illumina NovaSeq Platform

PE150

6 G/10 G/20 G

45 Aso faigaluega

Bioinformatics su'esu'ega

● Pulea lelei o fa'amaumauga mata'utia

● Fonotaga Metagenome

● Seti o kenera e le toe faia ma fa'amatalaga

● Su'esu'ega o ituaiga eseese

● Su'esu'ega o galuega fa'atupu kenera

● Su'esu'ega va'aiga

● Su'esu'ega fa'atasi e fa'atatau i mea fa'ata'ita'i

liuchengtu11

Fa'ata'ita'iga Mana'oga ma le Tu'uina atu

Fa'ata'ita'iga Mana'oga:

MoDNA extracts:

Ituaiga Fa'ata'ita'i

Aofaiga

Fa'atotonuga

mama

DNA extracts

> 30 ng

> 1 ng/μl

OD260/280= 1.6-2.5

Mo fa'ata'ita'iga tau si'osi'omaga:

Fa'ata'ita'iga ituaiga

Fautuaina faiga faataitai

Eleele

Aofa'i fa'ata'ita'iga: tusa.5 g;O mea maguma totoe e tatau ona aveese mai luga;Oli fasipepa lapopoa ma pasi i totonu ole faamama 2 mm;Aliquot faʻataʻitaʻiga i le sterile EP-tube poʻo le cyrotube mo le faʻaagaga.

Feces

Aofa'i fa'ata'ita'iga: tusa.5 g;Aoina ma fa'ata'ita'i fa'ata'ita'iga ile EP-tube po'o le cryotube mo le fa'aagaga.

I totonu o le manava

E mana'omia le fa'agaioiina o fa'ata'ita'iga i lalo ole tulaga aseptic.Fufulu aano o lo'o aoina ile PBS;Centrifuge le PBS ma fa'aputu le mea fa'afefe ile EP-tubes.

Sludge

Aofa'i fa'ata'ita'iga: tusa.5 g;Aoina ma fa'ata'ita'i fa'ata'ita'i palapala ile EP-tube po'o le cryotube mo le fa'aagaga

Vaivai

Mo faʻataʻitaʻiga e faʻatapulaʻa le aofaʻi o microbial, e pei o le paipa vai, vai vai, ma isi, Aʻoaʻo mai le itiiti ifo i le 1 L vai ma pasi ile 0.22 μm faamama e faʻatamaoaigaina ai microbial i luga ole membrane.Teu le pa'u i totonu o le faagaau mama.

Pa'u

'A'e ma le fa'aeteete le pa'u i le su'e fugala'au po'o le lau tipitipi ma tu'u i totonu ole faagaau mama.

Fautuaina Fa'ata'ita'iga Tu'uina atu

Fa'a'aisa fa'ata'ita'iga ile sua vai mo le 3-4 itula ma teu ile fa'asusu vai po'o le -80 tikeri ile fa'aagaga umi.Fa'ata'ita'iga felauaiga fa'atasi ai ma le aisa mago e mana'omia.

Auala o Galuega Au'aunaga

fa'ata'ita'iga tu'uina atu

Fa'ata'ita'iga tu'uina atu

Sauniuniga Faletusi

Fausia faletusi

Fa'asologa

Fa'asologa

Iloiloga fa'amaumauga

Iloiloga fa'amaumauga

A uma le fa'atauga Auaunaga

Au'aunaga pe a uma ona fa'atau


  • Muamua:
  • Sosoo ai:

  • 1.Histogram: Tufatufaina o ituaiga

    3

    2. Functional genes annotated to KEGG metabolic pathways

    4

    3. Heat fa'afanua: 'ese'ese galuega fa'avae e fa'atatau i le tele o gafa54.Circos of CARD antibiotic resistance genes

    6

    BMK mataupu

    Fa'ateleina o kenera fa'ama'i fa'ama'i fa'ama'i ma fa'ama'i fa'ama'i i luga ole eleele-togatogo fa'aauau

    Lolomiina:Tusitala o Mea Mata'utia, 2021

    Fuafuaga fa'asologa:

    Meafaitino: DNA fa'asoa mai vaega e fa o a'a togatogo fa'ata'ita'iga e feso'ota'i ma fa'ata'ita'iga: palapala e le'i totōina, rhizosphere, episphere ma vaega endosphere.
    Fa'avae: Illumina HiSeq 2500
    Sini: Metagenome
    16S rRNA gene V3-V4 itulagi

    I'uga autu

    Metagenomic sequencing ma metabarcoding profiling i luga o le eleele-a'a fa'aauau o togatogo sa fa'agasolo ina ia mafai ai ona su'esu'eina le fa'asalalauina o kenera fa'ama'i fa'ama'i (ARGs) mai le palapala i totonu o la'au.Fa'amatalaga Metagenomic na fa'aalia ai e 91.4% o kenera e tetee atu ai fa'ama'i fa'ama'i e masani ona iloa i vaega uma e fa o le palapala o lo'o ta'ua i luga, lea na fa'aalia ai le fa'aauau pea.16S rRNA amplicon sequencing gaosia 29,285 sequencing, fai ma sui o 346 ituaiga.O le tu'ufa'atasia ma le fa'avasegaina o ituaiga e ala i le fa'asologa o le amplicon, o lenei fa'asalalauga na maua e tuto'atasi mai le microbiota e feso'otai i a'a, peita'i, e mafai ona fa'afaigofieina e le telefoni feavea'i o elemene kenera.O lenei su'esu'ega na iloa ai le tafe mai o ARGs ma pathogens mai le palapala i totonu o la'au e ala i feso'ota'iga o le eleele-a'a.

    Fa'asinomaga

    Wang, C. , Hu, R. , Malosi, PJ , Zhuang, W. , & Shu, L. .(2020).Fa'ateleina o kenera fa'ama'i fa'ama'i ma fa'ama'i fa'ama'i i luga ole eleele-fa'aauau a'a togatogo.Tusitala o Mea Mata'utia, 408, 124985.

    maua se upusii

    Tusi lau savali iinei ma lafo mai ia i matou

    Auina mai lau savali ia i matou: