Fa'aitiitiga Fa'atusa o le Bisulfite Sequencing (RRBS)
Fa'aititia le Fa'asologa o le Bisulfite Sequencing (RRBS) ua tula'i mai e avea o se fa'aoga taugofie ma le lelei i le Whole Genome Bisulfite Sequencing (WGBS) i su'esu'ega DNA methylation. Aʻo tuʻuina atu e le WGBS faʻamatalaga manino e ala i le suʻesuʻeina o le genome atoa i le faʻavae e tasi, o lona tau maualuga e mafai ona avea ma mea faʻatapulaʻa. O le RRBS e faʻaitiitia ai lenei luʻitau e ala i le faʻavasegaina o se vaega sui o le genome. O lenei metotia e fa'alagolago i le fa'atamaoaigaina o le CpG i le motu-oa'oa e ala i le MspI cleavage soso'o ai ma le lapopo'a filifilia o vaega 200-500/600 bps. O le mea la lea, e na'o itulagi e lata ane i motu CpG e fa'asologa, a'o i latou e mamao atu motu CpG e le o aofia i le su'esu'ega. O lenei faagasologa, faʻatasi ma le bisulfite sequencing, e mafai ai ona maua le maualuga o le suʻesuʻeina o le DNA methylation, ma le faʻasologa o le auala, PE150, e taulaʻi faʻapitoa i pito o faʻapipiʻi nai lo le ogatotonu, faʻateleina le lelei o le methylation profiling. O le RRBS o se meafaigaluega taua tele e mafai ai ona taugofie suʻesuʻega DNA methylation ma faʻalauteleina le malamalama o masini epigenetic.
Vaega Auaunaga
● Manaomia se genome faasinomaga.
● Lambda DNA o loʻo faʻaaogaina e mataʻituina le lelei o le liua o le bisulfite.
● O lo'o mata'itūina fo'i le fa'amamaina o le MspI.
● fa'aluaina fa'ama'i fa'ama'i mo la'au fa'ata'ita'iga.
● Fa'asologa ile Illumina NovaSeq.
Tulaga Auaunaga
●Tau-lelei ma lelei Suiga i le WGBS: mafai ai ona faia le au'ili'iliga i se tau maualalo ma fa'ata'ita'iga mana'oga maualalo.
●Fa'atonu atoatoa:tu'uina atu auaunaga sili ona lelei e tasi mai le fa'ata'ita'iga fa'ata'ita'iga, fau faletusi, ma le fa'asologa i su'esu'ega o bioinformatics.
●Tomai Tele: fa'atasi ai ma poloketi fa'asologa o RRBS ua mae'a fa'amae'aina i le tele o ituaiga o ituaiga, BMKGENE e maua ai le sili atu ma le sefulu tausaga o le poto masani, o se 'au su'esu'e sili ona atamai, fa'amatalaga atoatoa, ma le lagolago sili ona lelei pe a mae'a fa'atau.
Fa'amatalaga Auaunaga
| Faletusi | Fuafuaga Fa'asologa | Fautuaina fa'amatalaga fa'amatalaga | Pulea lelei |
| MspI digested ma le Bisulfite faletusi | Illumina PE150 | 8 Gb | Q30 ≥ 85% Liuga ole bisulfite > 99% MspI lelei tipi > 95% |
Fa'ata'ita'iga Manaoga
| Fa'atotonuga (ng/µL) | Aofa'iga (µg) |
| |
| Genomic DNA | ≥ 30 | ≥ 1 | Fa'atapula'aina le fa'aleagaina po'o le fa'aleagaina |
Auala o Galuega Au'aunaga
Fa'ata'ita'iga tu'uina atu
Fausia faletusi
Fa'asologa
Iloiloga fa'amaumauga
Au'aunaga pe a uma ona fa'atau
E aofia ai le au'ili'iliga nei:
● Fa'atonuga lelei ole fa'asologa;
● Faafanua e faasino i le genome;
● Suʻesuʻeina o faʻavae methylated 5mC ma faʻamatalaga faʻailoga;
● Suʻesuʻega o le tufatufaina o le methylation ma faʻatusatusaga faʻataʻitaʻiga;
● Su'esu'ega o Itulagi Eseese Methylated (DMRs);
● Fa'amatalaga aoga o kenera e feso'ota'i ma DMRs.
Fa'atonuga lelei: fa'amama lelei (i fa'afanua genome)
Puleaina lelei: liua bisulfite (i le methylation faʻamatalaga faʻamatalaga)
Fa'afanua Methylation: 5mC methylation fa'asologa o genome-lautele
Fa'ata'ita'iga fa'atusa: Su'esu'ega Autu Vaega
Su'esu'ega o Itulagi Methylated Eseese (DMRs): fa'afanua vevela
Su'esu'e le alualu i luma o su'esu'ega na fa'afaigofieina e le BMKGene o le genome bisulfite sequencing services e ala i le aoina o fa'asalalauga.
Li, Z. et al. (2022) 'Faʻatonuga maualuga-faʻamaoni reprogramming i le Leydig-pei sela e ala i le CRISPR faʻagaoioia ma vaega paracrine',PNAS Nexus, 1(4). doi: 10.1093/PNASNEXUS/PGAC179.
Tian, H. et al. (2023) 'Genome-wide DNA methylation analysis of body composition in Chinese monozygotic masaga',European Journal of Clinical Investigation, 53(11), itulau. e14055. doi: 10.1111/ECI.14055.
Wu, Y. et al. (2022) 'DNA methylation ma le puimanava i le suilapalapa fua: o se suʻesuʻega faʻapitoa epigenome i masaga monozygotic Saina',Journal of Endocrinological Investigation, 45(12), itulau 2365–2376. doi: 10.1007/S40618-022-01878-4.
