Ulandelelwano lwe-bisulfite ye-genome epheleleyo (WGBS)
Le ndlela, esekelwe kunyango lwe-bisulfite lwe-DNA ukuze kuguqulwe i-cytosines ezingenamethylated zibe yi-uracil (C ukuya kwi-U), ngelixa ishiya i-cytosines ezinemethylated zingatshintshanga, imi njengendlela esemgangathweni ophezulu yokuhlola ngokunzulu i-methylation ye-DNA, ingakumbi indawo yesihlanu kwi-cytosine (5-mC), umlawuli obalulekileyo wokubonakaliswa kwezakhi zofuzo kunye nomsebenzi weseli.
Le ndlela inika isisombululo esisisiseko esinye, ivumela abaphandi ukuba baphande ngokupheleleyo i-methylome baze bafumanise iipatheni ze-methylation ezingaqhelekanga kwiisampuli. Ngokusebenzisa i-WGBS, izazinzulu zinokufumana ulwazi olungenakuthelekiswa nanto malunga neendawo ze-methylation ezijikelezileyo ze-genome, zibonelela ngokuqonda okucacileyo kweendlela ze-epigenetic ezisisiseko seenkqubo ezahlukeneyo zebhayoloji kunye nezifo.
Iimpawu zeNkonzo
● Ukulandelelana kwe-Illumina NovaSeq.
● Ifuna i-genome ebhekisa kwi-reference.
● I-DNA yeLambda yongezwa ukuze ijonge ukusebenza kakuhle kokuguqulwa kwe-bisulfite.
Iingenelo zeNkonzo
●Umgangatho weGolide woPhando lweMethylation yeDNA: Le teknoloji ivuthiweyo ichanekileyo kakhulu kwaye iyakwazi ukuphinda ivele.
●Ukugubungela okuBanzi kunye nesisombululo esisekwe kwisiseko esinye:Ukufunyanwa kweendawo ze-methylation kukwinqanaba le-genome ngokubanzi.
●Iqonga Elipheleleyo:Sinikezela ngenkonzo yokumisa yonke indawo ukusuka ekucwangcisweni kweesampuli, ukwakhiwa kwethala leencwadi, ukulandelelana ukuya kuhlalutyo lwe-bioinformatics.
●Ubuchule Obubanzi: Sineeprojekthi ezininzi ezigqityiweyo kwiintlobo ngeentlobo zeentlobo. I-BMKGENE izisa amava angaphezulu kweminyaka elishumi, iqela lohlalutyo elinobuchule obuphezulu, umxholo opheleleyo, kunye nenkxaso ebalaseleyo emva kokuthengisa.
●Amathuba okujoyina uHlalutyo lweTranscriptomics: Ukuvumela uhlalutyo oludibeneyo lwe-WGBS kunye nezinye iinkcukacha ze-omics ezifana ne-RNA-seq.
Iinkcukacha zeSampuli
| Ithala leeNcwadi | Icebo Lokulandelelanisa | Imveliso yedatha ecetyiswayo | Ulawulo lwemeko |
| Iphathwe ngeBisulfite | I-Illumina PE150 | Ubunzulu obungama-30x | Q30 ≥ 85% Ukuguqulwa kweBisulfite > 99% |
Iimfuno zeSampuli
| Uxinzelelo (ng/µL) | Isixa sisonke (µg) | Iimfuneko ezongezelelweyo | |
| I-DNA ye-genomic | ≥ 5 | ≥ 400 ng | Ukonakala okulinganiselweyo okanye ungcoliseko |
Ukuhamba komsebenzi wenkonzo
Ukuhanjiswa kwesampulu
Ukukhupha i-DNA
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Ukuhanjiswa kwedatha
Le nkonzo ibandakanya olu hlalutyo lulandelayo:
● Ulawulo lomgangatho wokulandelelana okungacwangciswanga;
● Ukumapha kwi-genome ebhekisa kwi-genome;
● Ukufunyanwa kweziseko ze-methylated ze-5mC;
● Uhlalutyo lokusasazwa kwe-methylation kunye nokuchazwa kwayo;
● Uhlalutyo lweeNdawo ezineMethylene eyahlukileyo (iiDMR);
● Inkcazo esebenzayo yezakhi zofuzo ezinxulumene ne-DMR.
Ukufunyanwa kwe-methylation ye-5mC: iintlobo zeendawo ezine-methylation
Imephu yeMethylation. Ukusasazwa kwe-genome ye-methylation ye-5mC ngokubanzi
Inkcazo yeendawo ezine-methyl eninzi
Iindawo ezineMethylene eyahlukileyo: iijini ezinxulumene nazo
IiNdawo ezineMethyletic ezahlukeneyo: inkcazelo yezakhi zofuzo ezinxulumene nazo (iGene Ontology)
Hlola inkqubela phambili yophando eququzelelwe ziinkonzo ze-BMKGene zokulandelelanisa i-genome bisulfite ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.
UFan, Y. et al. (2020) 'Uhlalutyo lweeprofayili ze-DNA methylation ngexesha lophuhliso lwemisipha yethambo legusha kusetyenziswa ulandelelwano lwe-bisulfite epheleleyo',I-BMC Genomics, 21(1), iphepha 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Iingxaki ezintsha ze-deoxyribonucleic acid methylation kubasebenzi abachaphazeleka yi-vinyl chloride',I-Toxicology kunye neMpilo yeShishini, 38(7), iphepha 377–388. doi: 10.1177/07482337221098600
UZuo, J. et al. (2020) 'Ubudlelwane phakathi kwe-genome methylation, amanqanaba e-RNAs ezingabhalisiyo ikhowudi, ii-mRNAs kunye nee-metabolites kwisiqhamo setamatato esivuthwayo',Ijenali yeZityalo, 103(3), iphepha 980-994. doi: 10.1111/TPJ.14778.
