Isku xigxiga genome bisulfite (WGBS)
Dhammaan Genome Bisulfite Sequencing (WGBS) waxay u taagan tahay habka heerka-dahabka ah ee sahaminta qoto dheer ee methyl DNA-da, gaar ahaan booska shanaad ee cytosine (5-mC), xakameynta muhiimka ah ee muujinta hiddaha iyo dhaqdhaqaaqa gacanta. Mabda'a ka hooseeya WGBS waxay ku lug leedahay daawaynta bisulfit, taasoo keenaysa beddelka cytosines-ka aan methylated ilaa uracil (C ilaa U), iyadoo ka tagaysa cytosines methylated oo aan isbeddelin. Farsamadani waxay bixisaa xallin hal-saldhig ah, taasoo u oggolaanaysa cilmi-baarayaasha inay si buuxda u baaraan methylome-ka oo ay ogaadaan qaababka methylation-ka ee aan caadiga ahayn ee la xiriira xaalado kala duwan, gaar ahaan kansarka. Shaqaalaysiinta WGBS, saynisyahannadu waxay heli karaan aragtiyo aan la qiyaasi karin oo ku saabsan muuqaallada methylation-ka ee genom-ballaaran, iyagoo siinaya faham qotodheer oo ku saabsan hababka epigenetic ee hoos yimaada hababka noole ee kala duwan iyo cudurrada.
Tilmaamaha Adeegga
● Wuxuu u baahan yahay genome tixraaceed.
● DNA-da Lambda ayaa lagu daraa si loo kormeero waxtarka beddelidda bisulfit.
● Taxanaha Illumina NovaSeq.
Faa'iidooyinka Adeegga
●Heerka Dahab ee Cilmi-baarista Methylation DNA: Tignoolajiyadan wax-ka-beddelka methylation ee qaan-gaadhka ah waxay leedahay saxnaan sare iyo taransiin wanaagsan.
●Daboolista Ballaaran iyo Qaraarka Saldhig-Kaliya:ogaanshaha goobaha methylation ee heerka genome-ballaaran.
●Madal Dhameystiran:Bixi adeeg aad u wanaagsan oo hal joogsi ah laga bilaabo habaynta muunada, dhismaha maktabadda, isku xigxiga ilaa falanqaynta bioinformatics.
●Khibrad Balaadhan: iyada oo mashaariicda isku xigxiga ee WGBS ay si guul leh u dhammaystireen noocyo kala duwan oo noocyo kala duwan ah, BMKGENE waxay keenaysaa toban sano oo waayo-aragnimo ah, koox falanqaynta xirfad sare leh, nuxur dhamaystiran, iyo taageero iib ah oo aad u fiican.
●Suurtagalnimada inaad ku biirto Falanqaynta Qoraal-qorista: u oggolaanaya falanqaynta isku dhafan ee WGBS xogta omics kale sida RNA-seq.
Tilmaamaha Tusaalaha
| Maktabadda | Istaraatiijiyada Taxanaha | Soo saarista xogta lagu taliyay | Xakamaynta tayada |
| Bisulfite waa la daaweeyay | Illumina PE150 | 30x qoto dheer | Q30 ≥ 85% Beddelka Bisulfite> 99% |
Shuruudaha Tusaalaha
| Xooga saarida (ng/µL) | Tirada guud (µg) | Shuruudaha dheeraadka ah | |
| DNA-da genomiga ah | ≥ 5 | ≥ 400 ng | Waxyeelo ama faddarayn xaddidan |
Socodka Shaqada Adeegga
Tusaalaha keenista
Soo saarista DNA
Dhismaha maktabadda
Isku xigxiga
Falanqaynta xogta
gaarsiinta xogta
Waxa ku jira falanqaynta soo socota:
● Xakamaynta tayada cayriinka ah;
● Khariidaynta tixraaca genome;
● Ogaanshaha 5mC saldhigyada methylated;
● Falanqaynta qaybinta methylation iyo sharraxaadda;
● Falanqaynta Gobollada Methylated ee Kala Duwan (DMRs);
● Sharaxaada hawleed ee hiddo-wadaha la xidhiidha DMR-yada.
5mC ogaanshaha methylation: noocyada goobaha methylated
Khariidadda Methylation 5mC methylation qaybinta genome-ballaaran
Faahfaahinta gobollada methylated aadka u sarreeya
Gobollada Methylated oo kala duwan: hidde-sideyaasha la xidhiidha
Gobollada Methylated oo kala duwan: sharraxaad hidde-sideyaasha la xidhiidha (Gene Ontology)
Sahami horumarka cilmi-baarista ee ay fududaysay BMKGene dhammaan adeegyada isku xigxiga genome bisulfite iyada oo loo marayo ururinta daabacaadyo la soo xulay.
Fan, Y. iyo al. (2020) 'Falanqaynta profiles DNA methylation inta lagu guda jiro horumarinta murqaha lafaha idaha iyadoo la adeegsanayo isku xigxiga bisulfite-genome-ga dhan',BMC Genomics, 21 (1), bogga 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. iyo al. (2022) 'Dhibaatooyinka methylation deoxyribonucleic acid novel ee shaqaalaha soo gaadhay chloride vinyl',Toxicology iyo Caafimaadka Warshadaha, 38(7), bogga 377-388. doi: 10.1177/07482337221098600
Zuo, J. iyo al. (2020) 'Xiriirka ka dhexeeya genome methylation, heerarka RNA-yada aan codeeyn, mRNAs iyo dheef-shiid kiimikaadka ku jira bislaanshaha miraha yaanyada',Wargeyska dhirta, 103(3), bogga 980–994. doi: 10.1111/TPJ.14778.
