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Iimveliso

Ulandelelwano lweMetagenomic (NGS)

I-Metagenome ibhekiselele kwingqokelela yemathiriyeli yemfuzo iyonke yoluntu oluxubeneyo lwezinto eziphilayo, ezifana ne-metagenome yokusingqongileyo, i-metagenome yabantu, njl njl.Ukulandelelana kwe-Metagenomic sisixhobo se-molecular esetyenziselwa ukuhlalutya izinto ezixubeneyo ze-genomic ezithathwe kwiisampuli zendalo, ezinikezela ngolwazi olucacileyo kwiintlobo zeentlobo zeentlobo kunye nobuninzi, ubume benani labantu, ubudlelwane be-phylogenetic, izakhi zofuzo ezisebenzayo kunye nenethiwekhi yokulungelelanisa kunye nezinto ezisingqongileyo.

Iqonga:Illumina NovaSeq6000


Iinkcukacha zeNkonzo

Iziphumo zeDemo

Into yokuphonononga

Iinkonzo eziluncedo

ØUkwahlukaniswa kunye nokulinywa simahla kwiprofayili yoluntu lwemicrobial

ØIsisombululo esiphezulu ekufumaneni iintlobo eziphantsi kobuninzi kwiisampuli zokusingqongileyo

ØIngcamango ye "meta-" idibanisa zonke iimpawu zebhayoloji kwinqanaba lokusebenza, inqanaba leentlobo kunye nenqanaba lemfuza, ebonisa umbono oguqukayo osondeleyo kwinyani.

ØI-BMK iqokelela amava amakhulu kwiintlobo zeesampulu ezahlukeneyo ezineisampulu ezingaphezulu kwe-10,000 ezicutshungulweyo.

IiNgcaciso zeNkonzo

UkulandelelanisaIqonga

Ithala leencwadi

Imveliso yedatha ecetyiswayo

Ixesha eliqikelelweyo lokujika

Illumina NovaSeq 6000

PE250

50K/100K/300K iithegi

Iintsuku ezingama-30

Uhlalutyo lwe-Bioinformatics

üUlawulo lomgangatho wedatha ekrwada

üIndibano yeMetagenome

üIseti yemfuza engafunekiyo kunye nenkcazo

üUhlalutyo lweentlobo ezahlukeneyo zeentlobo

üUhlalutyo lokwahluka komsebenzi wemfuza

üUhlalutyo lwamaqela

üUhlalutyo lombutho ngokuchasene nezinto zovavanyo

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IiMfuno zeSampulu kunye nokuhanjiswa

IiMfuno zeSampulu:

KubaIzicatshulwa zeDNA:

Uhlobo lweSampuli

Isixa

Ukugxila

Ubunyulu

Izicatshulwa zeDNA

> 30 ng

> 1 ng/μl

OD260/280= 1.6-2.5

Ngeesampulu zokusingqongileyo:

Uhlobo lwesampuli

Inkqubo yesampulu ecetyiswayo

Umhlaba

Isixa sesampulu: malunga.5 g;Into eseleyo ebunayo kufuneka isuswe kumphezulu;Gaya iinqununu ezinkulu kwaye udlule kwi-2 mm filter;Iisampulu ze-Aliquot kwityhubhu ye-EP okanye i-cyrotube yogcino.

Ilindle

Isixa sesampulu: malunga.5 g;Qokelela kwaye iisampulu ze-aliquot kwi-EP-tube engcolileyo okanye i-cryotube yokubhukisha.

Iziqulatho zamathumbu

Iisampuli kufuneka ziqwalaselwe phantsi kwemeko ye-aseptic.Hlamba izicubu eziqokelelweyo nge-PBS;I-Centrifuge i-PBS kwaye uqokelele i-precipitant kwi-EP-tubes.

Udaka

Isixa sesampulu: malunga.5 g;Qokelela kwaye isampuli ye-aliquot yodaka kwi-EP-tube engcolileyo okanye i-cryotube yokugcinelwa

Amanzi

Kwisampulu enesixa esilinganiselweyo se-microbial, njengamanzi epompo, amanzi omthombo, njl., Qokelela ubuncinci be-1 L yamanzi kwaye udlule kwi-0.22 μm yokucoca ukutyebisa i-microbial kwi-membrane.Gcina inwebu kwityhubhu enyumba.

Ulusu

Khulula ngononophelo umphezulu wolusu ngeswab yomqhaphu engcolileyo okanye ngencakuba yotyando kwaye uyibeke kwityhubhu engenanto.

Unikezelo lweSampuli eCetyisiweyo

Umkhenkce iisampulu kulwelo nitrogen iiyure 3-4 kwaye ugcine kulwelo nitrogen okanye -80 degree ukuya ugcino ixesha elide.Isampulu yokuthumela ngenqanawe eyomileyo iyafuneka.

Ukuhamba komsebenzi weNkonzo

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Ukuhanjiswa kwesampuli

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Ukwakhiwa kwethala leencwadi

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Ukulandelelanisa

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Uhlalutyo lwedatha

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Iinkonzo emva kokuthengisa


  • Ngaphambili:
  • Okulandelayo:

  • 1.I-Histogram: Ukuhanjiswa kweentlobo

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    2.Ufuzo olusebenzayo oluchazwe kwiindlela ze-metabolic ze-KEGG

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    3.Imephu yobushushu: Imisebenzi eyohlukileyo esekelwe kubuninzi bemfuza5I-4.I-Circos ye-CARD i-antibiotic resistance genenes

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    Ityala le-BMK

    Ukuxhaphaka kofuzo lokuxhathisa amayeza okubulala iintsholongwane kunye neentsholongwane zebhaktiriya ecaleni kwengcambu yommango

    Ipapashiwe:Ijenali yeZixhobo ezinobungozi, ngo-2021

    Iqhinga lokulandelelanisa:

    Izinto: Izicatshulwa ze-DNA zamaqhekeza amane eengcambu zemangrove ezinxulumene neesampuli: umhlaba ongatyalwanga, irhizosphere, i-episphere kunye ne-endosphere compartments
    Iqonga: Illumina HiSeq 2500
    Ujoliso: iMetagenome
    16S rRNA gene V3-V4 ummandla

    Iziphumo eziphambili

    Ulandelelwano lwe-Metagenomic kunye ne-metabarcoding profiling kwi-root-root continuum ye-mangrove saplings yacutshungulwa ukuze kufundwe ukusasazwa kofuzo lwe-antibiotic resistance (ARGs) ukusuka emhlabeni ukuya kwizityalo.Idatha ye-Metagenomic ibonise ukuba i-91.4% yejene yokumelana namayeza okubulala iintsholongwane aye achongwa ngokuqhelekileyo kuwo onke amacandelo omhlaba akhankanywe ngasentla, abonisa ifashoni eqhubekayo.Ulandelelwano lwe-amplicon ye-16S rRNA yenze i-29,285 yokulandelelana, emele iintlobo ze-346.Ukudibanisa neentlobo zeprofayili ngokulandelelana kwe-amplicon, oku kusasazwa kwafunyaniswa kuzimeleyo kwi-microbiota ehambelana neengcambu, nangona kunjalo, inokuququzelelwa yi-mobile ye-genetic elements.Olu phononongo luchonge ukuhamba kwee-ARG kunye neentsholongwane ezibangela izifo ezisuka emhlabeni zisiya kwizityalo ngokudityaniswa kweengcambu zomhlaba.

    Isalathiso

    Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).Ukuxhaphaka kwejini yokuxhathisa amayeza okubulala iintsholongwane kunye neentsholongwane ezibangela iintsholongwane ecaleni kwengcambu yommango.Ijenali yeZixhobo ezinobungozi, 408, 124985.

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