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条形 isibhengezo-03

Imikhiqizo

I-Reduced Representation Bisulfite Sequencing (RRBS)

Ucwaningo lwe-DNA methylation belulokhu luyisihloko esishisayo ocwaningweni lwezifo, futhi luhlobene kakhulu nezakhi zofuzo kanye nezici ze-pheno-typic.I-RRBS iyindlela enembile, ephumelelayo neyongayo yocwaningo lwe-DNA methylation.Ukunothiswa komgqugquzeli kanye nezifunda zesiqhingi ze-CpG nge-enzymatic cleavage (Msp I), kuhlanganiswe nokulandelana kwe-Bisulfite, kunikeza ukutholwa kwe-DNA methylation yokuxazulula okuphezulu.

Ipulatifomu: I-Illumina NovaSeq Platform


Imininingwane Yesevisi

Umphumela wedemo

Imininingwane Yesevisi

Ingxenyekazi: NovaSeq platform, PE150

Uhlobo lwelabhulali: 200-400bp faka umtapo wezincwadi we-DNA we-bisulfite

Isu lokulandelanisa: Okubhangqiwe-Isiphetho esingu-150 bp

Ncoma okuphumayo kwedatha: 10 Gb idatha/isampula eluhlaza

Izidingo Zesampula

Ubuningi be-DNA: ≥ 2ug

Ukugxila kwe-DNA:≥ 20ng/μl.

Ukuhlanzeka: OD260/280 = 1.8 kuya ku-2.0 ngaphandle kokucekelwa phansi noma ukungcola kwe-RNA

Ukuhlaziywa kwe-Bioinformatics

流程图 羽4-03

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:

  • a.Izibalo zokuqondanisa i-genome yereferensi

    Izibalo zokufundwa kwemephu
    isithombe3

    Sizibalo zokujula kokulandelana kanye nokufakwa kwesayithi C

    isithombe4

    Aukufakwa okuqoqiwe

    isithombe5

    Dukusebenza kahle kokugaya kwesampula ngayinye

    isithombe6

    b.Ukuthola indawo ye-methylation

    Uhlu lwamazinga e-methylation endaweni ye-C

    isithombe7

    BS ukuguqulwa

    isithombe8 isithombe9

    Mi-ethylated cytosine
    isithombe10

    Ukusatshalaliswa kwezinga le-methylation
    isithombe11

    c.Imephu ye-methylation

    Genome-wide
    isithombe12

    Ginto eyodwa

    isithombe13

    d.Ukuqhathaniswa kwamazinga e-methylation phakathi kwamasampuli
    isithombe14

    PAirwise Ukuhlobana kwe-CG methylation
    isithombe15

    PI-CA-CG methylation
    isithombe16

    Cukuvuvukala kwe-CG methylation
    isithombe17

    d.Ukuhlaziywa kwe-methylation okuhlukile

    Sizibalo ze-DMR

    isithombe18

    Ukusabalalisa ubude be-DMR
    isithombe19

    Hidla imephu
    isithombe20

    DUkusatshalaliswa kwe-MR endaweni okuhlosiwe
    Isithombe21

    thola isaphulelo

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