Gbogbo genome bisulfite sequencing(WGBS)
Gbogbo Genome Bisulfite Sequencing (WGBS) duro bi ilana boṣewa goolu fun iwadii ijinle ti DNA methylation, ni pataki ipo karun ni cytosine (5-mC), olutọsọna pataki ti ikosile pupọ ati iṣẹ ṣiṣe cellular. Ilana ti o wa labẹ WGBS pẹlu itọju bisulfite, ti nfa iyipada ti awọn cytosines ti ko ni methylated si uracil (C si U), lakoko ti o nlọ awọn cytosine methylated ko yipada. Ilana yii nfunni ni ipinnu ipilẹ-ẹyọkan, gbigba awọn oniwadi laaye lati ṣe iwadii ni kikun lori methylome ati ṣii awọn ilana methylation ajeji ti o ni nkan ṣe pẹlu awọn ipo pupọ, ni pataki akàn. Nipa lilo WGBS, awọn onimo ijinlẹ sayensi le jèrè awọn oye ti ko lẹgbẹ sinu awọn ala-ilẹ methylation jakejado-genome, n pese oye ti o ni oye ti awọn ọna ṣiṣe epigenetic ti o wa labẹ awọn ilana ati awọn aarun onirũru.
Awọn ẹya ara ẹrọ Iṣẹ
● Nbeere genome itọkasi kan.
● Lambda DNA ti wa ni afikun lati ṣe atẹle ṣiṣe iyipada bisulfite.
● Sequencing lori Illumina NovaSeq.
Awọn anfani Iṣẹ
●Iwọn goolu fun Iwadi Methylation DNA: Imọ-ẹrọ iyipada methylation ogbo yii ni iṣedede giga ati atunṣe to dara.
●Ibora Fife ati Ipinnu Ipilẹ Kanṣo:wiwa ti awọn aaye methylation ni ipele jiini jakejado.
●Pipe Platform:pese ọkan-Duro o tayọ iṣẹ lati awọn ayẹwo processing, ìkàwé ikole, sequencing to bioinformatics onínọmbà.
●Amoye nla: pẹlu awọn iṣẹ akanṣe WGBS ti pari ni aṣeyọri kọja awọn oriṣiriṣi oriṣiriṣi oriṣiriṣi, BMKGENE mu iriri iriri ọdun mẹwa lọ, ẹgbẹ itupalẹ oye ti o ga julọ, akoonu okeerẹ, ati atilẹyin ti o dara julọ lẹhin-tita.
●O ṣeeṣe lati Darapọ mọ pẹlu Itupalẹ Transcriptomics: gbigba fun itupalẹ iṣọpọ ti WGBS pẹlu awọn data omics miiran bii RNA-seq.
Apeere pato
| Ile-ikawe | Ilana titele | Ijade data ti a ṣe iṣeduro | Iṣakoso didara |
| Bisulfite ṣe itọju | Imọlẹ PE150 | 30x ijinle | Q30 ≥ 85% Bisulfite iyipada> 99% |
Awọn ibeere apẹẹrẹ
| Ifojusi (ng/µL) | Apapọ iye (µg) | Awọn ibeere afikun | |
| DNA jinomiki | ≥ 5 | ≥ 400 ng | Idibajẹ to lopin tabi ibajẹ |
Sisan Iṣẹ Iṣẹ
Ifijiṣẹ apẹẹrẹ
DNA isediwon
Ikole ìkàwé
Titele
Itupalẹ data
Ifijiṣẹ data
Pẹlu itupale wọnyi:
● Aise sequencing didara iṣakoso;
● Ìyàwòrán sí ìtọ́ka-ara-ara;
● Wiwa ti awọn ipilẹ methylated 5mC;
● Ayẹwo ti pinpin methylation ati akọsilẹ;
● Ayẹwo ti Awọn Agbegbe Methylated Iyatọ (DMRs);
● Alaye iṣẹ-ṣiṣe ti awọn Jiini ti o ni nkan ṣe pẹlu awọn DMR.
5mC methylation erin: orisi ti methylated ojula
Methylation maapu. 5mC methylation jiini-jakejado pinpin
Alaye ti awọn agbegbe methylated giga
Awọn Agbegbe Methylated Iyatọ: Awọn Jiini ti o somọ
Awọn agbegbe Methylated Iyatọ: asọye ti awọn Jiini ti o somọ (Gene Ontology)
Ṣawakiri awọn ilọsiwaju iwadii ti o ni irọrun nipasẹ BMKGene gbogbo awọn iṣẹ itọsẹ bisulfite genome nipasẹ ikojọpọ ti awọn atẹjade.
Fan, Y. et al. (2020) 'Onínọmbà ti awọn profaili methylation DNA lakoko idagbasoke iṣan egungun ti agutan nipa lilo gbogbo-ẹda bisulfite bisulfite-genome',BMC Genomics, 21 (1), ojú ìwé 1–15. doi: 10.1186 / S12864-020-6751-5.
Zhao, X. et al. (2022) 'Aratuntun deoxyribonucleic acid methylation perturbations ninu awọn oṣiṣẹ ti o farahan si fainali kiloraidi',Toxicology ati Ilera Ile-iṣẹ, 38 (7), ojú ìwé 377–388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Awọn ibatan laarin genome methylation, awọn ipele ti awọn RNA ti kii ṣe ifaminsi, mRNAs ati awọn metabolites ni eso tomati ti o pọn',Iwe akosile ohun ọgbin, 103 (3), oju ewe 980–994. doi: 10.1111 / TPJ.14778.
