Ukulandelana kwe-bisulfite ye-genome ephelele (WGBS)
Le ndlela, esekelwe ekwelashweni kwe-bisulfite kwe-DNA ukuze kuguqulwe ama-cytosine angena-methylated abe yi-uracil (C kuya ku-U), kuyilapho ishiya ama-cytosine angena-methylated engashintshiwe, iyindlela ejwayelekile yokuhlola ngokujulile i-methylation ye-DNA, ikakhulukazi indawo yesihlanu ku-cytosine (5-mC), umlawuli obalulekile wokubonakaliswa kwezakhi zofuzo kanye nomsebenzi wamaseli.
Le ndlela inikeza isisombululo sesisekelo esisodwa, okuvumela abacwaningi ukuthi baphenye ngokuphelele i-methylome futhi bathole amaphethini e-methylation angavamile ngaphakathi kwamasampula. Ngokusebenzisa i-WGBS, ososayensi bangathola ukuqonda okungenakuqhathaniswa mayelana nezindawo ze-methylation ezibanzi ze-genome, okuhlinzeka ngokuqonda okuningiliziwe kwezindlela ze-epigenetic ezisekela izinqubo nezifo ezahlukahlukene zezinto eziphilayo.
Izici Zesevisi
● Ukulandelana kwezithombe ku-Illumina NovaSeq.
● Kudinga i-genome ebhekisela kuyo.
● I-DNA ye-Lambda yengezwa ukuze kuqashwe ukusebenza kahle kokuguqulwa kwe-bisulfite.
Izinzuzo Zesevisi
●Izinga Legolide Locwaningo Lwe-DNA Methylation: Lobu buchwepheshe obuvuthiwe bunokunemba okuphezulu kanye nokuphindaphindeka okuhle.
●Ukumbozwa Okubanzi kanye nesisombululo sesisekelo esisodwa:Ukutholwa kwezindawo ze-methylation kusezingeni elibanzi le-genome.
●Ipulatifomu Ephelele:Sinikeza isevisi yokuma okukodwa kusukela ekucubungulweni kwamasampula, ukwakhiwa komtapo wolwazi, ukulandelana kuya ekuhlaziyweni kwe-bioinformatics.
●Ubuchwepheshe Obubanzi: Sinezinhlobonhlobo zamaphrojekthi aqediwe ezinhlobonhlobo zezinhlobo. I-BMKGENE iletha ulwazi olungaphezu kweminyaka eyishumi, ithimba lokuhlaziya elinekhono eliphezulu, okuqukethwe okuphelele, kanye nokusekelwa okuhle kakhulu ngemuva kokuthengisa.
●Kungenzeka Ukujoyina Ukuhlaziywa Kwe-Transcriptomics: Ukuvumela ukuhlaziywa okuhlanganisiwe kwe-WGBS neminye idatha ye-omics njenge-RNA-seq.
Imininingwane Yesampula
| Umtapo Wolwazi | Isu lokulandelana kwezigaba | Ukukhishwa kwedatha okunconyiwe | Ikhwalithi yokulawula |
| Ilashwe nge-bisulfite | I-Illumina PE150 | Ukujula okungu-30x | Q30 ≥ 85% Ukuguqulwa kwe-Bisulfite > 99% |
Izidingo Zesampula
| Ukugxila (ng/µL) | Inani eliphelele (ng) | Izidingo ezengeziwe | |
| I-DNA ye-Genomic | ≥ 1.25 | ≥ 100 | Ukonakala noma ukungcola okulinganiselwe |
Ukugeleza Komsebenzi Wesevisi
Ukulethwa kwesampula
Ukukhishwa kwe-DNA
Ukwakhiwa komtapo wolwazi
Ukulandelana
Ukuhlaziywa kwedatha
Ukulethwa kwedatha
Le nsizakalo ifaka phakathi ukuhlaziywa okulandelayo:
● Ukulawulwa kwekhwalithi yokulandelana okungahluziwe;
● Ukuhlela i-genome ebhekisele kuyo;
● Ukutholwa kwezisekelo ze-methylated ze-5mC;
● Ukuhlaziywa kokusatshalaliswa kwe-methylation kanye nokuchazwa kwayo;
● Ukuhlaziywa Kwezindawo Ezine-Methylated Ehlukile (ama-DMR);
● Isichasiselo esisebenzayo sezakhi zofuzo ezihlotshaniswa nama-DMR.
Ukutholwa kwe-methylation ye-5mC: izinhlobo zezindawo ezine-methylation
Imephu ye-Methylation. Ukusatshalaliswa kwe-genome ye-methylation ye-5mC
Isichasiselo sezindawo ezine-methyl eningi
Izifunda Ezine-Methylene Ehlukile: izakhi zofuzo ezihlobene
Izifunda Ezine-Methylated Ngokuhlukile: isichasiselo sezakhi zofuzo ezihlobene (i-Gene Ontology)
Hlola intuthuko yocwaningo eyenziwe yizinsizakalo zokulandelana kwe-genome bisulfite ze-BMKGene ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
UFan, Y. et al. (2020) 'Ukuhlaziywa kwamaphrofayili e-DNA methylation ngesikhathi sokuthuthukiswa kwemisipha yamathambo ezimvu kusetshenziswa ukulandelana kwe-bisulfite ye-genome ephelele',I-BMC Genomics, 21(1), amakhasi 1-15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Ukuphazamiseka okusha kwe-deoxyribonucleic acid methylation kubasebenzi abachayeke ku-vinyl chloride',I-Toxicology kanye Nempilo Yezimboni, 38(7), amakhasi 377-388. doi: 10.1177/07482337221098600
UZuo, J. et al. (2020) 'Ubudlelwano phakathi kwe-genome methylation, amazinga ama-RNA angabhalisi ikhodi, ama-mRNA kanye nama-metabolites esithelweni sikatamatisi esivuthwayo',Ijenali Yezitshalo, 103(3), amakhasi 980-994. doi: 10.1111/TPJ.14778.
