Hoʻemi ʻia ka hoʻohālikelike Bisulfite Sequencing (RRBS)
Ua puka mai ka Reduced Representation Bisulfite Sequencing (RRBS) ma ke ʻano he kumu kūʻai kūpono a maikaʻi hoʻi i ka Whole Genome Bisulfite Sequencing (WGBS) i ka noiʻi methylation DNA. ʻOiai hāʻawi ʻo WGBS i nā ʻike kikoʻī ma o ka nānā ʻana i ka genome holoʻokoʻa ma ka hoʻonā kumu hoʻokahi, hiki i kāna kumukūʻai kiʻekiʻe ke lilo i mea palena. Hoʻopau maikaʻi ʻo RRBS i kēia pilikia ma ke koho ʻana i kahi ʻāpana o ka genome. Ke hilinaʻi nei kēia ʻano hana i ka hoʻonui ʻia ʻana o nā ʻāina waiwai nui o ka mokupuni ʻo CpG e ka MspI cleavage a ukali ʻia e ka koho nui o nā ʻāpana 200-500/600 bps. No laila, ʻo nā ʻāina kokoke wale i nā mokupuni CpG i hoʻokaʻawale ʻia, ʻoiai ʻo nā mea me nā mokupuni CpG mamao e kāpae ʻia mai ka nānā ʻana. ʻO kēia kaʻina hana, i hui pū ʻia me ka bisulfite sequencing, e ʻae i ka ʻike kiʻekiʻe o ka methylation DNA, a me ke ala hoʻonohonoho, PE150, e nānā pono i nā wēlau o nā mea hoʻokomo ma mua o ka waena, e hoʻonui ana i ka pono o ka methylation profiling. He mea waiwai nui ka RRBS e hiki ai i ka noiʻi DNA methylation ke kumu kūʻai a hoʻonui i ka ʻike o nā mīkini epigenetic.
Nā hiʻohiʻona lawelawe
● Pono ka genome kuhikuhi.
● Hoʻohana ʻia ʻo Lambda DNA e nānā i ka maikaʻi o ka hoʻololi ʻana i ka bisulfite.
● Mākaukau ʻia ka pono o ka digestion MspI.
● ʻO ka hoʻoheheʻe ʻana i ka enzyme pālua no nā laʻana meakanu.
● Ke kaʻina hana ma Illumina NovaSeq.
Pono lawelawe
●ʻO ke kumu kūʻai kūpono a maikaʻi hoʻi i ka WGBS: hiki ke hoʻokō ʻia ka nānā ʻana ma ke kumu kūʻai haʻahaʻa a me nā koi haʻahaʻa haʻahaʻa.
●Papahana piha:hāʻawi i ka lawelawe maikaʻi loa mai ka hoʻoili ʻana i nā laʻana, kūkulu hale waihona puke, a me ka hoʻonohonoho ʻana i ka loiloi bioinformatics.
●ʻIke Nui: me RRBS sequencing projects i hoʻopau maikaʻi ʻia ma waena o nā ʻano ʻano like ʻole, lawe mai ʻo BMKGENE i nā makahiki he ʻumi o ka ʻike, kahi hui loiloi mākaukau loa, ʻike piha, a me ke kākoʻo maikaʻi loa ma hope o ke kūʻai aku.
Nā kikoʻī lawelawe
| Hale Waihona Puke | Hoʻolālā Sequencing | Hoʻopuka ʻia ka ʻikepili i manaʻo ʻia | Ka hoomalu maikai |
| MspI digested a me Bisulfite hale waihona puke | Illumina PE150 | 8 Gb | Q30 ≥ 85% Hoʻololi bisulfite > 99% MspI ʻoki pono ʻana > 95% |
Nā Koina Laʻana
| Hoʻopaʻa (ng/µL) | huina nui (µg) |
| |
| DNA Genomic | ≥ 30 | ≥ 1 | Hoʻohaʻahaʻa palena ʻole a hoʻohaumia paha |
Holo Hana Hana
Hāʻawi laʻana
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Nā lawelawe ma hope o ke kūʻai aku
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 a me ka ʻike kumu;
● Ka nānā 'ana i ka māhele methylation a me ka ho'ohālikelike ho'ohālike;
● Nānā i nā ʻāpana ʻokoʻa methylated (DMRs);
● Hōʻike kikoʻī o nā genes pili i nā DMR.
Ka mana maikaʻi: ka hoʻoemi ʻana (ma ka palapala ʻāina genome)
Ka mana maikaʻi: hoʻololi bisulfite (ma ka ʻike ʻike methylation)
Palapala methylation: 5mC methylation genome-ākea mahele
Ka hoʻohālikelike laʻana: ʻIke ʻĀpana Haʻawina
Ka nānā 'ana i nā 'āpana Methylated Differentially (DMRs): heatmap
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.
Li, Z. et al. (2022) 'Ka hoʻoponopono hou ʻana i ka hilinaʻi kiʻekiʻe i loko o nā cell e like me Leydig e ka CRISPR activation a me nā mea paracrine',PNAS Nexus, 1(4). hana: 10.1093/PNASNEXUS/PGAC179.
Tian, H. et al. (2023) 'Genome-wide DNA methylation analysis of body composition in Chinese monozygotic twins',European Journal of Clinical Investigation, 53(11), p. e14055. doi: 10.1111/ECI.14055.
Wu, Y. et al. (2022) 'DNA methylation a me ka waist-to-hip ratio: kahi epigenome-wide association study in Chinese monozygotic twins',Nūpepa o ka Endocrinological Investigation, 45(12), pp. 2365–2376. doi: 10.1007/S40618-022-01878-4.
