BMKCloud Log in
条形 isibhengezo-03

Imikhiqizo

16S/18S/ITS Amplicon Sequencing-PacBio

I-subunit ku-16S kanye ne-18S rRNA equkethe zombili izifunda ezilondolozwe kakhulu neziguquguqukayo iwuphawu lweminwe oluphelele lwemolekyuli yokuhlonza i-prokaryotic kanye ne-eukaryotic organisms.Ngokusebenzisa ithuba lokulandelanisa, lawa ma-amplicon angaqondiswa ngokusekelwe ezingxenyeni ezilondoloziwe futhi izifunda eziguquguqukayo zingabonakaliswa ngokugcwele ukuhlonza i-microbial enomthelela ocwaningweni oluhlanganisa ukuhlaziywa kwe-microbial diversity, taxonomy, phylogeny, njll. I-molecule ye-Single-real-time(SMRT) ) ukulandelana kweplathifomu ye-PacBio kwenza kutholwe ukufundwa okude okunembe kakhulu, okungahlanganisa ama-amplicon anobude obugcwele (cishe u-1.5 KB).Ukubuka okubanzi kwendima yofuzo kuthuthukise kakhulu ukulungiswa kwesichasiselo sezinhlobo kubhaktheriya noma emphakathini wesikhunta.

Inkundla:I-PacBio Sequel II


Imininingwane Yesevisi

Imiphumela yedemo

Ucwaningo lwesigameko

Izinzuzo Zesevisi

1

● Ukufunda isikhathi eside kuveza ukulandelana kobude obugcwele be-16S/18S/ITS

● Ukushaya kwesisekelo okunembe kakhulu okulandelanayo kwemodi ye-PacBio CCS

● Ukulungiswa kwezinga lezinhlobo kusichasiselo se-OTU/ASV

● Ukugeleza kokuhlaziya kwe-QIIME2 kwakamuva nokuhlaziya okuhlukahlukene ngokuya kusizindalwazi, isichasiselo, i-OTU/ASV.

● Kusebenza ezifundweni ezihlukahlukene zomphakathi ezincane

● I-BMK inokuhlangenwe nakho okubanzi okunamasampula angaphezu kuka-100,000/ngonyaka, emboza inhlabathi, amanzi, igesi, udaka, indle, amathumbu, isikhumba, umhluzi obilayo, izinambuzane, izitshalo, njll.

● I-BMKCloud iqondise ukuhunyushwa kwedatha equkethe amathuluzi okuhlaziya omuntu angama-45

Imininingwane Yesevisi

UkulandelanisaInkundla

Umtapowolwazi

Idatha enconyiwe

Isikhathi sokushintsha

I-PacBio Sequel II

Insimbi ye-SMRT

5K/10K/20K Omaka

44 Izinsuku zokusebenza

I-Bioinformatics ihlaziya

● Ukulawulwa kwekhwalithi yedatha eluhlaza

● I-OTU clustering/De-noise(ASV)

● Isichasiselo se-OTU

● Ukuhlukahluka kwe-alpha

● Ukuhlukahluka kwe-Beta

● Ukuhlaziya kwamaqembu

● Ukuhlaziywa kwenhlangano ngokumelene nezici zokuhlola

● Isibikezelo sofuzo lomsebenzi

16sPacbio

Izidingo Zesampula Nokulethwa

Izimfuneko zesampula:

NgobaIzingcaphuno ze-DNA:

Uhlobo lwesampula

Inani

Ukugxila

Ubumsulwa

Izingcaphuno ze-DNA

> 1 μg

> 20 ng/μl

OD260/280= 1.6-2.5

Ngamasampuli emvelo:

Uhlobo lwesampula

Inqubo yokusampula enconyiwe

Umhlabathi

Inani lesampula: cishe.5 g;Izinto ezisele ezibunile zidinga ukususwa phezulu;Gaya izingcezu ezinkulu futhi udlule ngesihlungi esingu-2 mm;Amasampuli e-Aliquot ku-EP-tube eyinyumba noma i-cyrotube ukuze abhukhe.

Indle

Inani lesampula: cishe.5 g;Qoqa futhi amasampula e-aliquot ku-EP-tube eyinyumba noma i-cryotube ukuze ubhukhe.

Okuqukethwe kwamathumbu

Amasampula adinga ukucutshungulwa ngaphansi kwesimo se-aseptic.Geza izicubu eziqoqiwe nge-PBS;I-Centrifuge i-PBS futhi uqoqe i-precipitant kumashubhu e-EP.

Udaka

Inani lesampula: cishe.5 g;Qoqa futhi isampula ye-aliquot sludge ku-EP-tube eyinyumba noma i-cryotube ukuze ubhukhe

Amanzi

Ukuze uthole isampula enenani elilinganiselwe le-microbial, njengamanzi kampompi, amanzi omthombo, njll., Qoqa okungenani amanzi angu-1 L futhi udlule kusihlungi esingu-0.22 μm ukuze unothise i-microbial kulwelwesi.Gcina ulwelwesi kushubhu oyinyumba.

Isikhumba

Hlanza ngokucophelela indawo yesikhumba ngeswabhu kakotini oyinyumba noma i-blade yokuhlinza bese uyibeka eshubhuni eliyinyumba.

Ukulethwa Kwesampula Okunconyiwe

Friza amasampula ku-nitrogen ewuketshezi amahora angu-3-4 futhi uwagcine ku-nitrogen ewuketshezi noma -80 degree ekubhukeni isikhathi eside.Kudingeka isampula yokuthunyelwa ngeqhwa elomile.

Ukugeleza Komsebenzi Wesevisi

ukulethwa kwesampula

Ukulethwa kwesampula

Ukulungiselela Umtapowolwazi

Ukwakhiwa komtapo wolwazi

Ukulandelanisa

Ukulandelanisa

Ukuhlaziywa kwedatha

Ukuhlaziywa kwedatha

Ngemva kokudayiswa Services

Amasevisi angemuva kokuthengisa


  • Okwedlule:
  • Olandelayo:

  • 1.Isilinganiso sesichasiselo se-V3+V4(Illumina)-based microbial community profileing vs. Full-length (PacBio)-based profileing.
    (Idatha yamaphrojekthi angu-30 akhethwe ngokungahleliwe afakwe kwizibalo)

    3

    2.Isilinganiso sesichasiselo sokulandelana kwe-amplicon yobude obugcwele ezingeni lezinhlobo zezinhlobo zesampuli ezihlukene

    4

    3.Ukusatshalaliswa kwezinhlobo zezilwane

    5
    4.Isihlahla se-Phylogenetic

    6

    Icala le-BMK

    Ukuchayeka kwe-arsenic kubangela ukulimala kwesithiyo samathumbu kanye nokwenza kusebenze i-axis yesibindi se-gut okuholela ekuvuvukeni kanye ne-pyroptosis yesibindi kumadada.

    Kushicilelwe:Isayensi Yendawo Yonke,2021

    Isu lokulandelanisa:

    Amasampuli: Ukulawula vs 8 mg/kg iqembu eliveziwe le-ATO
    Ilandelanisa isivuno sedatha: 102,583 ukulandelana kwe-CCS eluhlaza sekukonke
    Ukulawula: 54,518 ± 747 CCS esebenzayo
    I-ATO-eveziwe : 45,050 ± 1675 i-CCS esebenzayo

    Imiphumela esemqoka

    Ukuhlukahluka kwe-Alpha:Ukuchayeka kwe-ATO kushintshe kakhulu ukunotha kwamagciwane emathunjini kanye nokwehluka kwamadada.

    Ukuhlaziywa kwe-Metastats:
    Ezingeni le-phylum: I-bacterial phyla engu-2 itholakala kuphela emaqenjini okulawula
    Ezingeni lohlobo: Izinhlobo eziyisi-6 zitholwe zihluke kakhulu ngobuningi obuhlobene
    Ngokwezinga lezinhlobo zezilwane: izinhlobo ezingama-36 zikhonjwe sezizonke, eziyisi-6 zazo zihluke kakhulu ngobuningi bokuphila.

    Ireferensi

    Thingholm, LB , et al."Abantu abakhuluphele abanesifo sikashukela sohlobo 2 nabangenaso babonisa amandla nokusebenza okuhlukile kwe-Gut Microbial Functional."I-Cell Host & Microbe26.2(2019).

    thola isaphulelo

    Bhala umlayezo wakho lapha futhi usithumelele wona

    Thumela umlayezo wakho kithi: