I-genome BIsul FI Te tempking (i-WGBS)
Ukuhlelwa kabusha kwe-genome bisulfite yonke kumi njengendlela ye-Gold-Standard Methology yokuhlola okujulile kwe-DNA methylation, ikakhulukazi isikhundla sesihlanu ku-cytosine (5-Mc), umlawuli obaluleke kakhulu we-gene expression kanye nomsebenzi wamaselula. Isisekelo se-WGBS ebesindayo kubandakanya ukwelashwa kwe-Bisulfite, efaka ukuguqulwa kwama-cytosines angafakwanga ku-Uracil (c ku-U), ngenkathi ushiya ama-cytosines we-methylated engashintshiwe. Le ndlela inikeza ukulungiswa kwesisekelo esisodwa, okuvumela abacwaningi ukuthi baphenye ngokugcwele i-methylome futhi baveze amaphethini we-methylation angejwayelekile ahambisana nemibandela ehlukahlukene, ikakhulukazi umdlavuza. Ngokuqasha ama-WGBS, ososayensi bangathola imininingwane engafani ne-genome-wind methylation andscapes, enikeza ukuqonda okungenzeka kwama-epigenetic izindlela ezihlukene zemvelo nezifo.
Izici Zensizakalo
● kudinga i-genome yereferensi.
● I-Lambda DNA yengezwa ukuqapha ukusebenza kahle kwe-Bisulfite.
● Ukulandelana ku-Illina Novaseq.
Izinzuzo zensizakalo
●I-Gold Standard ye-DNA methylation ucwaningo: Lokhu kuguqulwa kwe-methylation methylation ukuguqulwa kwe-technology kunokunemba okuphezulu kanye nokuzala okuhle.
●Ukuvalwa okubanzi kanye nesixazululo esiyisisekelo esisodwa:ukutholwa kwamasayithi we-methylation ezingeni elibanzi.
●Isikhulumi Esiphelele:Nikeza ngezinsizakalo zokuma okuhle kakhulu kusuka ekucutshungweni kwesampula, ukwakhiwa kwetabhulali, ukulandelana kokuhlaziywa kwe-bioinformatics.
●Ubuchwepheshe obubanzi: Ngemiklamo yokulandelana kwe-WGBS iphothule ngempumelelo ezinhlobonhlobo ezahlukahlukene zezinhlobo, i-BMKGENE iletha ngaphezulu kweshumi leminyaka leminyaka, iqembu lokuhlaziya elinamakhono kakhulu, okuqukethwe okuphelele, nokusekelwa okuhle kokuthengisa okungcono.
●Amathuba okujoyina nokuhlaziywa kwe-transcriptics: Ukuvumela ukuhlaziywa okuhlanganisiwe kwama-WGBs nge eminye imininingwane ye-OMICS efana ne-RNA-SEQ.
Ukucaciswa kwesampula
| Indlu ehlala izincwadi | Ukuhlehlisa Isu | Okunconyiwe kwedatha okunconyiwe | Ikhwalithi yokulawula |
| I-Bisulfite yaphathwa | I-Illumina PE150 | Ukujula kwe-30x | Q30 ≥ 85% Ukuguqulwa kwe-Bisulfite> 99% |
Izidingo zesampula
| Ukuhlushwa (ng / μl) | Inani eliphelele (μg) | Izidingo ezingeziwe | |
| I-genomic DNA | ≥ 5 | ≥ 400 ng | Ukuwohloka okulinganiselwe noma ukungcoliswa |
Ukugeleza komsebenzi wesevisi
Ukulethwa kwesampula
Ukukhishwa kwe-DNA
Ukwakhiwa Kwelabhulali
Ukulandelana
Ukuhlaziywa kwedatha
Ukulethwa Kwedatha
Kubandakanya ukuhlaziywa okulandelayo:
● Ukulawulwa kwekhwalithi okuluhlaza okulula;
● Ukuthola imephu ukuze ubheke i-genome;
● Ukutholwa kwezisekelo ze-5mc methyled;
● Ukuhlaziywa kokusatshalaliswa kwe-methylation kanye nesichasiselo;
● Ukuhlaziywa kwezifunda ezihlukile ze-methylated (DMR);
● Isichasiselo esisebenzayo sezakhi zofuzo ezihambisana ne-DMRS.
Ukutholwa kwe-5mc methylation: Izinhlobo zamasayithi afakiwe
Imephu ye-methylation. I-5mc methylation genome-genome-genome
Isichasiselo sezifunda eziphakeme kakhulu
Izifunda ezihlukile ze-methylated: izinhlobo ezihambisana nazo
Izifunda ezihlukile ze-methylated: Isichasiselo sezakhiso ezihambisanayo (i-gene ontology)
Hlola intuthuko yocwaningo ebika izinsizakalo zokulandelana kwezinsizakalo zeBmkgene zeBorkgene's Genome Bisulfite Services ngeqoqo elikhethiwe lokushicilelwa.
Fan, y et al. (2020) 'Ukuhlaziywa kwamaphrofayili we-DNA methylation ngesikhathi sokuthuthukiswa kwemisipha yezimvu usebenzisa ukulandelana kwe-bisulfite yonke',BMC genomics, 21 (1), kk. 1-15. Doi: 10.11186 / S12864-020-6751-5.
UZhao, X. et al. .I-TOXICICOLOGY NEMPILO YEMPILO, 38 (7), PP. 377-388. doi: 10.11177 / 07482337221098600
UZuo, J. et al. (2020) 'ubudlelwano phakathi kwe-genome methylation, amazinga ama-rnas angewona amakhodi, ama-mrnas kanye nama-metabolites ngezithelo zamatamatisi ezivuthiwe',Iphephabhuku lezitshalo, 103 (3), PP. 980-994. DOI: 10.11111 / TPJ.14778.
