ʻO ke kaʻina hana bisulfite genome holoʻokoʻa (WGBS)
Ke kū nei ʻo Whole Genome Bisulfite Sequencing (WGBS) ma ke ʻano he kumu maʻamau gula no ka ʻimi hohonu ʻana i ka DNA methylation, ʻo ia hoʻi ke kūlana ʻelima i ka cytosine (5-mC), kahi mea hoʻoponopono pivotal o ka hōʻike gene a me ka hana kelepona. ʻO ke kumu o ka WGBS e pili ana i ka lāʻau bisulfite, e hoʻohuli i ka hoʻololi ʻana o nā cytosines unmethylated i uracil (C to U), ʻoiai e waiho ana i nā cytosines methylated i hoʻololi ʻole ʻia. Hāʻawi kēia ʻenehana i ka hoʻonā kumu hoʻokahi, e ʻae i nā mea noiʻi e noiʻi piha i ka methylome a wehe i nā ʻano methylation kūlohelohe e pili ana i nā kūlana like ʻole, ʻo ia hoʻi ka maʻi kanesa. Ma ka hoʻohana ʻana iā WGBS, hiki i nā ʻepekema ke loaʻa nā ʻike like ʻole i nā ʻāina methylation ākea genome, e hāʻawi ana i ka ʻike nuanced o nā mīkini epigenetic e hoʻokumu i nā kaʻina hana olaola like ʻole a me nā maʻi.
Nā hiʻohiʻona lawelawe
● Pono ka genome kuhikuhi.
● Hoʻohui ʻia ʻo Lambda DNA e nānā i ka maikaʻi o ka hoʻololi ʻana i ka bisulfite.
● Ke kaʻina hana ma Illumina NovaSeq.
Pono lawelawe
●Kūlana gula no ka ʻimi noiʻi DNA Methylation: He kiʻekiʻe kiʻekiʻe ka pololei a me ka reproducibility maikaʻi kēia ʻenehana hoʻololi hoʻololi methylation.
●Ka uhi ākea a me ka ʻōlelo hoʻoholo hoʻokahi:ka ʻike ʻana i nā pūnaewele methylation ma ka pae genome-ākea.
●Papahana piha:hāʻawi i ka lawelawe maikaʻi loa mai ka hoʻoheheʻe ʻana i ka laʻana, ke kūkulu hale waihona puke, ka hoʻonohonoho ʻana i ka loiloi bioinformatics.
●ʻIke Nui: me nā papahana hoʻonohonoho WGBS i hoʻopau maikaʻi ʻia ma waena o nā ʻano ʻano like ʻole, lawe mai ʻo BMKGENE i kahi ʻumi makahiki o ka ʻike, kahi hui loiloi mākaukau loa, ʻike piha, a me ke kākoʻo maikaʻi loa ma hope o ke kūʻai ʻana.
●Hiki ke hui pū me ka Transcriptomic Analysis: e 'ae ana i ka ho'opili 'ana o WGBS me nā 'ikepili omics e like me RNA-seq.
Hōʻike Laʻana
| Hale Waihona Puke | Hoʻolālā Sequencing | Hoʻopuka ʻia ka ʻikepili i manaʻo ʻia | Ka hoomalu maikai |
| Hana ʻia ka bisulfite | Illumina PE150 | 30x hohonu | Q30 ≥ 85% Hoʻololi bisulfite > 99% |
Nā Koina Laʻana
| Hoʻopaʻa (ng/µL) | huina nui (µg) | Nā koi hou aku | |
| DNA Genomic | ≥ 5 | ≥ 400 ng | Hoʻohaʻahaʻa palena ʻole a hoʻohaumia paha |
Holo Hana Hana
Hāʻawi laʻana
Ka unuhi ʻana o DNA
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Hāʻawi ʻikepili
Loaʻa i kēia ka nānā ʻana:
● Ka hoʻomalu ʻana i ka maikaʻi o ke kaʻina ʻana;
● Ka palapala ʻāina no ka kuhikuhi ʻana i ka genome;
● ʻIke i nā kumu methylated 5mC;
● Ka nānā 'ana i ka māhele methylation a me ka hō'ike 'ana;
● Nānā i nā ʻāpana ʻokoʻa methylated (DMRs);
● Hōʻike kikoʻī o nā genes pili i nā DMR.
5mC methylation ʻike: ʻano o nā pūnaewele methylated
Palapala methylation. 5mC methylation genome-ākea mahele
Hōʻike no nā wahi methylated kiʻekiʻe
Nā ʻāpana Methylated ʻokoʻa: nā genes pili
Nā ʻāpana Methylated ʻokoʻa: hōʻike no nā genes pili (Gene Ontology)
E ʻimi i nā holomua noiʻi i hoʻomaʻamaʻa ʻia e BMKGene nā lawelawe hoʻokaʻina genome bisulfite holoʻokoʻa ma o ka hōʻiliʻili ʻana o nā puke.
Fan, Y. et al. (2020) 'Ka nānā ʻana i nā profiles methylation DNA i ka wā o ka hoʻomohala ʻana i ka ʻiʻo o ka ʻiʻo hipa me ka hoʻohana ʻana i ka sequencing bisulfite holoʻokoʻa-genome',BMC Genomics, 21(1), pp. 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Novel deoxyribonucleic acid methylation perturbations i nā limahana i hōʻike ʻia i ka vinyl chloride',Hoʻonaʻauao a me ke Ola Hana Hana, 38(7), pp. 377–388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Ka pilina ma waena o ka genome methylation, nā pae o nā RNA non-coding, mRNA a me nā metabolites i ka oʻo ʻana i ka hua ʻōmato',Ka Nupepa Mea kanu, 103(3), pp. 980–994. doi: 10.1111/TPJ.14778.
