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条形 ibhanile-03

Iimveliso

16S/18S/ITS Amplicon Sequencing-PacBio

Iyunithi ekwi-16S kunye ne-18S rRNA equlathe yomibini imimandla egcinwe kakhulu kunye ne-hyper-variable printing yeminwe egqibeleleyo yemolekyuli yokuchongwa kweprokaryotic kunye ne-eukaryotic organisms.Ukuthatha ithuba lolandelelwano, ezi amplicon zinokujoliswa ngokusekwe kumacandelo agciniweyo kwaye imimandla eguquguqukayo ephezulu inokubonakaliswa ngokupheleleyo ukuchongwa kwemicrobial enegalelo kwizifundo ezibandakanya uhlalutyo lwentlupheko yemicrobial, taxonomy, phylogeny, njl. ) ulandelelwano lweqonga lePacBio lwenza ukuba kufumaneke ufundo olude oluchaneke kakhulu, olunokugubungela i-amplicon yobude obupheleleyo (malunga ne-1.5 KB).Imbono ebanzi yendawo yemfuza yasiphucula kakhulu isisombululo kwingcaciso yeentlobo kwiibhaktiriya okanye kuluntu lomngundo.

Iqonga:I-PacBio Sequel II


Iinkcukacha zeNkonzo

Iziphumo zeDemo

Into yokuphonononga

Iinkonzo eziluncedo

1

● Ufundo olude luveza ulandelelwano olupheleleyo lwe-16S/18S/ITS

● Isiseko esichanekileyo sokufowuna kunye nolandelelwano lwemodi ye-PacBio CCS

● Isisombululo senqanaba leentlobo kwinkcazo ye-OTU/ASV

● I-QIIME2 yamva nje yokuhlalutya ukuhamba nohlalutyo olwahlukeneyo ngokwemigaqo yogcino-lwazi, inkcazo, i-OTU/ASV.

● Isetyenzwa kwizifundo zoluntu ezahlukeneyo zemicrobial

● I-BMK inamava abanzi ngeesampulu ezingaphezulu kwe-100,000 ngonyaka, ezigquma umhlaba, amanzi, igesi, udaka, ilindle, amathumbu, ulusu, umhluzi wokubila, izinambuzane, izityalo, njl.

● I-BMKCloud iququzelele ukutolikwa kwedatha equlethe i-45 yezixhobo zokuhlalutya zomntu

IiNgcaciso zeNkonzo

UkulandelelanisaIqonga

Ithala leencwadi

Idatha ecetyiswayo

Ixesha lokuguqula

I-PacBio Sequel II

SMRT-intsimbi

5K/10K/20K iithegi

44 Iintsuku zokusebenza

Uhlalutyo lwe-Bioinformatics

● Ulawulo lomgangatho wedatha ekrwada

● I-OTU clustering/De-noise(ASV)

● Inkcazo ye-OTU

● Ukwahluka kweAlpha

● Ukwahluka kweBeta

● Uhlalutyo lwamaqela ngamaqela

● Uhlalutyo lombutho ngokuchasene nemiba yovavanyo

● Uqikelelo lwemfuza yokusebenza

16sPacbio

IiMfuno zeSampulu kunye nokuhanjiswa

IiMfuno zeSampulu:

KubaIzicatshulwa zeDNA:

Uhlobo lweSampuli

Isixa

Ukugxininisa

Ubunyulu

Izicatshulwa zeDNA

> 1 μg

> 20 ng/μl

OD260/280= 1.6-2.5

Ngeesampuli 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 kwi-EP-tube engcolileyo 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 ye-sludge 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 iblade 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

ukuhanjiswa kwesampuli

Ukuhanjiswa kwesampuli

ULungiselelo lweThala leeNcwadi

Ukwakhiwa kwethala leencwadi

Ukulandelelanisa

Ukulandelelanisa

Uhlalutyo lwedatha

Uhlalutyo lwedatha

Emva kweeNkonzo zentengiso

Iinkonzo emva kokuthengisa


  • Ngaphambili:
  • Okulandelayo:

  • I-1.Izinga le-Annotation ye-V3 + V4 (Illumina)-based microbial community profiling vs. Full-length (PacBio)-based profiling.
    (Idatha yeeprojekthi ezingama-30 ezichongwe ngokungacwangciswanga ziye zafakwa kwizibalo)

    3

    2.Ireyithi yenkcazo yolandelelwano lweamplicon yobude obupheleleyo kwinqanaba leentlobo ezahlukeneyo zesampulu

    4

    3.Ukuhanjiswa kweentlobo

    5
    4.Umthi wePhylogenetic

    6

    Ityala le-BMK

    Ukuvezwa kwe-arsenic kubangela umonakalo othintela amathumbu kunye nokulandelwa kokusebenza kwe-gut-liver axis ekhokelela ekudumbeni kunye ne-pyroptosis yesibindi kumadada.

    Ipapashiwe:Inzululwazi yeNdawo yonke,2021

    Iqhinga lokulandelelanisa:

    Iisampulu: Ukulawula vs 8 mg / kg ATO evezwe iqela
    Ukulandelelanisa isivuno sedatha: 102,583 ulandelelwano lwe-CCS ekrwada iyonke
    Ulawulo: 54,518 ± 747 CCS esebenzayo
    I-ATO-eveziweyo : 45,050 ± 1675 i-CCS esebenzayo

    Iziphumo eziphambili

    Ukwahluka kweAlpha:Ukuvezwa kwe-ATO kutshintshe kakhulu ubutyebi be-intestinal microbial kunye nokwahluka kwamadada.

    Uhlalutyo lwe-Metastats:
    Kwinqanaba le-phylum: I-2 ibhaktheriya phyla ifunyenwe kuphela kumaqela olawulo
    Kwinqanaba le-genus: I-6 genera ifunyenwe yahluke kakhulu kubuninzi obunxulumene
    Kwinqanaba leentlobo: Iindidi ezingama-36 zachongwa zizonke, kunye ne-6 yazo yahluke kakhulu kubuninzi be-relavive

    Isalathiso

    Thingholm, LB , et al."Abantu abatyebileyo abanesifo seswekile kunye nabangenaso iSifo seswekile sesi-2 babonisa isakhono esiSebenzayo seThumbu leMicrobial."Umamkeli weSeli kunye neMicrobe26.2(2019).

    fumana isicatshulwa

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