Raupapatanga bisulfite genome (WGBS)
Ko te Katoa o te Genome Bisulfite Sequencing (WGBS) te tikanga koura-paerewa mo te torotoro hohonu mo te methylation DNA, inaa ko te tuarima o te tuunga i roto i te cytosine (5-mC), he kaiwhakahaere tino nui o te whakaputa ira me te mahi pūkoro. Ko te maataapono kei raro i te WGBS ko te maimoatanga bisulfite, e whakaawe ana i te huringa o nga cytosines unmethylated ki te uracil (C ki U), i te wa e waiho ana nga cytosines methylated kia kore e rereke. Ko tenei tikanga e tuku ana i te whakataunga turanga-kotahi, ka taea e nga kairangahau te tirotiro whanui i te methylome me te hura i nga tauira methylation rerekee e pa ana ki nga momo ahuatanga, ara te mate pukupuku. Ma te whakamahi i te WGBS, ka taea e nga kaiputaiao te mohio ki nga whenua methylation puta noa i te ira-ira, me te maarama ki nga tikanga epigenetic e whai ana i nga momo tikanga koiora me nga mate.
Nga waahanga ratonga
● Me whai ira tohutoro.
● Ka taapirihia te DNA Lambda ki te aro turuki i te pai o te hurihanga bisulfite.
● Whakaraupapa i runga i te Illumina NovaSeq.
Painga Ratonga
●Paerewa koura mo te Rangahau Methylation DNA: Ko tenei hangarau tukatuka whakawhiti methylation pakeke he tino tika me te pai o te reproducibility.
●Kapi Whānui me te Whakatau turanga-kotahi:te kitenga o nga waahi methylation i te taumata ira-whanui.
●Paerewa Whakaoti:whakarato ratonga pai kotahi-mutu mai i te tukatuka tauira, te hanga whare pukapuka, te raupapatanga ki te tātari bioinformatics.
●Tohunga Nui: me nga kaupapa raupapa WGBS kua oti pai puta noa i nga momo momo kanorau, ka kawea mai e BMKGENE neke atu i te tekau tau o te wheako, he roopu tātari tino mohio, he korero matawhānui, me te tautoko pai i muri i te hokonga.
●Ka taea te hono atu ki te Transcriptomic Analysis: tuku mo te tātaritanga whakauru o WGBS me etahi atu raraunga omics penei i te RNA-seq.
Tauira Tauira
| Whare Pukapuka | Rautaki Raupapa | Putanga raraunga e taunaki ana | Mana kounga |
| Ko te bisulfite te rongoa | Ilumina PE150 | 30x te hohonu | Q30≥85% Te hurihanga bisulfite >99% |
Nga Whakaritenga Tauira
| Kukū (ng/µL) | Te tapeke (µg) | Nga whakaritenga taapiri | |
| DNA genomic | ≥30 | ≥3 | He iti te whakaheke, te poke ranei |
Rere Mahi Ratonga
Te tuku tauira
tangohanga DNA
Te hanga whare pukapuka
Whakaraupapa
Te tātari raraunga
Te tuku raraunga
Kei roto ko te tātaritanga e whai ake nei:
● Raw sequencing mana kounga;
● Mahere ki te tohutoro iraira;
● Te kitenga o nga turanga methylated 5mC;
● Te tātaritanga o te tohatoha methylation me te tuhi korero;
● Tātaritanga o ngā Rohe Methylated Differentially (DMRs);
● Tuhituhi mahi o nga ira e hono ana ki nga DMR.
Te kitenga methylation 5mC: nga momo waahi methylated
Mapi Methylation. 5mC methylation tohatoha ira-whanui
He korero mo nga rohe tino methylated
Rohe Methylated Rerekē: ira whai pānga
Rohe Methylated Differentially: tuhipoka o nga ira e hono ana (Gene Ontology)
Torotoro i nga mahi rangahau i whakahaerea e nga ratonga whakaraupapa bisulfite genome katoa a BMKGene na roto i te kohinga whakaputanga.
Fan, Y. et al. (2020) 'Te wetewete i nga korero mo te methylation DNA i te wa o te whakawhanaketanga o nga uaua kōiwi hipi ma te whakamahi i te raupapa bisulfite-genome katoa',BMC Genomics, 21(1), api 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Nga raruraru methylation waikawa deoxyribonucleic hou i roto i nga kaimahi e pa ana ki te vinyl chloride',Toxicology me te Hauora Ahumahi, 38(7), api 377–388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Nga hononga i waenga i te methylation genome, nga taumata o nga RNA kore-whakawaehere, nga mRNA me nga metabolites i roto i te maoa o nga hua tōmato',Te Tuhituhi tipu, 103(3), wh.980–994. doi: 10.1111/TPJ.14778.
