I-Reduced Representation Bisulfite Sequencing (RRBS)
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
Ukugeleza Komsebenzi Wesevisi
Ukulethwa kwesampula
Ukwakhiwa komtapo wolwazi
Ukulandelanisa
Ukuhlaziywa kwedatha
Amasevisi angemuva kokuthengisa
a.Izibalo zokuqondanisa i-genome yereferensi
Izibalo zokufundwa kwemephu
Sizibalo zokujula kokulandelana kanye nokufakwa kwesayithi C
Aukufakwa okuqoqiwe
Dukusebenza kahle kokugaya kwesampula ngayinye
b.Ukuthola indawo ye-methylation
Uhlu lwamazinga e-methylation endaweni ye-C
BS ukuguqulwa
Mi-ethylated cytosine
Ukusatshalaliswa kwezinga le-methylation
c.Imephu ye-methylation
Genome-wide
Ginto eyodwa
d.Ukuqhathaniswa kwamazinga e-methylation phakathi kwamasampuli
PAirwise Ukuhlobana kwe-CG methylation
PI-CA-CG methylation
Cukuvuvukala kwe-CG methylation
d.Ukuhlaziywa kwe-methylation okuhlukile
Sizibalo ze-DMR
Ukusabalalisa ubude be-DMR
Hidla imephu
DUkusatshalaliswa kwe-MR endaweni okuhlosiwe
