Whole genome bisulfite sequencing(WGBS)
Whole Genome Bisulfite Sequencing (WGBS) e eme e le mokhoa o tloaelehileng oa khauta bakeng sa ho fuputsa ka botebo DNA methylation, haholo-holo boemo ba bohlano ho cytosine (5-mC), molaoli oa bohlokoa oa polelo ea lefutso le tšebetso ea liselefouno. Molao-motheo o thehiloeng ho WGBS o kenyelletsa phekolo ea bisulfite, e susumetsang phetoho ea li-cytosine tse se nang methylated ho uracil (C ho ea ho U), ha li ntse li siea li-cytosine tsa methylated li sa fetohe. Mokhoa ona o fana ka tharollo ea motheo o le mong, o lumellang bafuputsi ho etsa lipatlisiso ka botlalo ka methylome le ho sibolla mekhoa e sa tloaelehang ea methylation e amanang le maemo a fapaneng, haholo mofetše. Ka ho sebelisa WGBS, bo-ramahlale ba ka fumana leseli le ke keng la bapisoa mabapi le genome-wide methylation landscapes, ho fana ka kutloisiso e fapaneng ea mekhoa ea epigenetic e bakang lits'ebetso le mafu a fapaneng a tlhaho.
Likarolo tsa Tšebeletso
● E hloka genome ea referense.
● Lambda DNA e eketsoa ho shebella katleho ea phetoho ea bisulfite.
● Sequencing ho Illumina NovaSeq.
Melemo ea Tšebeletso
●Khauta Standard bakeng sa DNA Methylation Research: Theknoloji ena e tsoetseng pele ea ho fetola methylation e na le ho nepahala ho phahameng le ho ikatisa hantle.
●Phatlalatso e Pharalletseng le Qeto e le 'ngoe:ho fumanoa ha libaka tsa methylation boemong ba genome-wide.
●Sethala se Felletseng:fana ka ts'ebeletso e ntle ea sebaka se le seng ho tloha ts'ebetsong ea sampole, kaho ea laeborari, tatellano ea tlhahlobo ea bioinformatics.
●Tsebo e Pharalletseng: ka merero ea tatellano ea WGBS e phethiloeng ka katleho ho mefuta e fapaneng ea mefuta e fapaneng, BMKGENE e tlisa boiphihlelo bo fetang lilemo tse leshome, sehlopha sa tlhahlobo se nang le boiphihlelo, litaba tse felletseng, le tšehetso e ntle haholo ea kamora thekiso.
●Monyetla oa ho Kopana le Tlhahlobo ea Transcriptomics: ho lumella tlhahlobo e kopaneng ea WGBS le lintlha tse ling tsa omics joalo ka RNA-seq.
Litlhaloso tsa Mohlala
| Laebrari | Leano la Tatelano | Tlhahiso ea data e khothalelitsoeng | Taolo ea boleng |
| Pheko ea Bisulfite | Illumina PE150 | 30x botebo | Q30 ≥ 85% Phetoho ea Bisulfite > 99% |
Litlhoko tsa Mohlala
| Khokahano (ng/µL) | Kakaretso (µg) | Litlhoko tse eketsehileng | |
| Genomic DNA | ≥5 | ≥ 400 ng | Phokotso e fokolang kapa tshilafatso |
Phallo ea Mosebetsi oa Ts'ebeletso
Mohlala oa thomello
Ho ntšoa ha DNA
Kaho ea laebrari
Tatelano
Tlhahlobo ea lintlha
Phano ea data
E kenyelletsa tlhahlobo e latelang:
● Taolo ea boleng ba tatellano e tala;
● Ho etsa 'mapa oa litšupiso tsa genome;
● Ho fumanoa ha 5mC methylated bases;
● Tlhahlobo ea kabo ea methylation le tlhaloso;
● Tlhahlobo ea Libaka tse sa tšoaneng tsa Methylated (DMRs);
● Tlhaloso e sebetsang ea liphatsa tsa lefutso tse amanang le li-DMR.
Ho lemoha 5mC methylation: mefuta ea libaka tsa methylated
'Mapa oa methylation. 5mC methylation genome-wide distribution
Tlhaloso ea libaka tse nang le methylated haholo
Libaka tse fapaneng tsa Methylated: liphatsa tsa lefutso tse amanang
Libaka tse fapaneng tsa Methylated: tlhaloso ea liphatsa tsa lefutso tse amanang le tsona (Gene Ontology)
Lekola tsoelo-pele ea lipatlisiso e tsamaisoang ke litšebeletso tsa tatellano ea liphatsa tsa lefutso tsa BMKGene ka pokello e hlophisitsoeng ea lingoliloeng.
Fan, Y. et al. (2020) 'Tlhahlobo ea lintlha tsa DNA methylation nakong ea nts'etsopele ea mesifa ea linku e sebelisa tatellano ea genome bisulfite e felletseng',BMC Genomics, 21(1), leqepheng la 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Novel deoxyribonucleic acid methylation perturbations ho basebetsi ba pepesehetseng vinyl chloride',Toxicology le Bophelo bo Botle ba Liindasteri, 38(7), leqepheng la 377-388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Likamano lipakeng tsa genome methylation, maemo a li-RNA tse sa ngolisoeng, li-mRNA le metabolites ho tholoana ea tamati e butsoitseng',The Plant Journal, 103(3), leqepheng la 980-994. doi: 10.1111/TPJ.14778.
