Tag nrho genome bisul fi trective (wgbs)
Tag nrho genome bisulfite excuring (wgbs) sawv raws li kev tshawb nrhiav kub ntawm cov methylation, tshwj xeeb ntawm kev tswj hwm thib tsib ntawm cov noob qhia thiab cov kev ua haujlwm ntawm cev. Lub hauv paus ntsiab lus hauv qab wgbs cuam tshuam kev kho mob los ntawm kev hloov pauv ntawm unmethylated cytosines mus rau uracil (c rau koj), thaum tawm ntawm metylated cytosines unchangated tsis hloov. Cov txheej txheem no muaj kev daws teeb meem ib leeg, pub cov kws tshawb nrhiav kom nkag siab txog methylome thiab nthuav tawm cov qauv methylation txuam nrog ntau yam kev mob, tsis muaj qog nqaij hlav. Los ntawm kev ua haujlwm WGB, cov kws tshawb fawb tuaj yeem tau txais kev nkag siab zoo rau hauv genome-dav methylation ua hauj lwm zoo li cov txheej txheem epigenetic uas underlie cov txheej txheem sib txawv thiab cov kab mob.
Cov Nta
● Yuav tsum tau siv Genome.
● Lambda DNA yog ntxiv rau saib xyuas bisulfite hloov zuj zus kev ua tau zoo.
● Kev sib txuas ntawm Illina Novase.
Kev Pabcuam Zoo
●Cov qauv kub rau DNA Methylation kev tshawb nrhiav: Qhov kev paub me me methylation hloov siab thev naus laus zis muaj qhov siab dua thiab zoo replucibility.
●Kev tiv thaiv dav thiab ib leeg-daws teeb meem:Nrhiav tau ntawm cov chaw methylation ntawm qib genome-dav.
●Ua tiav platform:Muab ib qho kev pabcuam zoo tshaj plaws los ntawm cov qauv ua qauv, kev tsim kho tsev qiv ntawv, ua ntu zus, kev txheeb xyuas cov khoom lag luam.
●Kev txawj ntse dav: Nrog WGB nrho cov phiaj xwm ua tiav tau ua tiav ntau hom, bmkgene coj dhau kev paub, cov ntsiab lus tsom xam, thiab kev muag khoom tom qab zoo nkauj.
●Tau koom nrog nrog Transcriptomics Kev Tshawb Xyuas: Tso cai rau kev txheeb xyuas cov khoom sib xyaw ntawm WGB nrog lwm cov ntaub ntawv Omics xws li RNA-Seq.
Qauv Cov Lus Qhia
| Lub tsev qiv ntawv | Seitencing Lub Tswv Yim | Pom zoo cov ntaub ntawv tso zis | Kev Tswj Xyuas Zoo |
| Bisulfite kho | ICATINA PE150 | 30x tob | Q30 ≥ 85% Bisulfite Conterersion> 99% |
Qauv Yuav Tsum Tau
| Concentration (ng / μl) | Tag nrho cov nyiaj (μg) | Ntxiv Yuav Tsum Tau | |
| Genomic DNA | ≥ 5 | ≥ 400 ng | Txwv tsawg tsawg lossis paug tsis huv |
Kev Pabcuam Khiav
Qauv Xa
DNA extraction
Kev Siv Hauv Tsev Qiv Ntawv
Ze
Cov Ntaub Ntawv Kev Ntsuam Xyuas
Cov Ntaub Ntawv Xa
Xws li kev tsom xam hauv qab no:
● Rutrens kev tswj hwm zoo;
● Mapping genome;
● Nrhiav kom tau ntawm 5mC methylated hauv paus;
Kev tshuaj xyuas ntawm methylation faib thiab cov lus qhia;
Kev Ntsuam Xyuas ntawm cov cheeb tsam methylated (DMRs);
● Cov haujlwm ua haujlwm tshaj tawm cov noob muaj feem cuam tshuam nrog DMRS.
5MC Methylation Nrhiav tau: Hom chaw methylated
Methylation Daim Ntawv Qhia. 5mc methylation genome-dav faib
Annotation ntawm cov methylated thaj chaw
Cov methylated thaj tsam: Koom haum noob caj noob
Cov methylated thaj tsam: Tshaj tawm ntawm cov noob neej (gene oftology)
Tshawb cov kev tshawb fawb txog kev tshawb fawb los ntawm BMKGene tag nrho Genome Bisulfite ua ntu zus los ntawm kev sau cov ntaub ntawv curated.
Ntxuam, Y. li al. (2020) Kev txheeb xyuas ntawm DNA methylation profiles thaum cov menyuam yaus txoj kev loj hlob ntawm cov leeg pob txha pob txha pob txha pob txha los siv tag nrho-genome bisulfite sequencing ',BMC Genomics, 21 (1), pp. 1-15. DOI: 10.1186 / s12864-020-6751-5.
Zhao, X. li al. (2022) 'Tus tshiab deoxyribonucleic acid methylation penururations nyob rau hauv cov neeg ua haujlwm raug rau Vinyl Chloride',Toxicology thiab Kev Noj Qab Haus Huv, 38 (7), pp. 377-388. DOI: 10.1177 / 07482337222298600
Zuo, J. li al. (2020) 'Kev sib raug zoo ntawm cov methylation genome, qib tsis-coding rnas, modnas thiab metabolites nyob rau ripening txiv hmab txiv ntoo ",Cov phau ntawv xov xwm cog, 103 (3), PP. 980-994. DOI: 10.1111 / tPJ.14778.
