Fa'asologa atoa o le genome bisulfite (WGBS)
O le Genome Bisulfite Sequencing (WGBS) o loʻo tu e pei o le faʻataʻitaʻiga auro mo suʻesuʻega loloto o le DNA methylation, aemaise o le tulaga lona lima i le cytosine (5-mC), o se faʻatonuga taua o le faʻaaliga o kene ma gaioiga feaveaʻi. O le mataupu faavae o le WGBS e aofia ai togafitiga bisulfite, faʻaosofia le liua o cytosines unmethylated i le uracil (C i U), ae tuʻu le methylated cytosines e le suia. O lenei metotia e ofoina atu ai le iugafono e tasi, e mafai ai e tagata suʻesuʻe ona suʻesuʻe atoatoa le methylome ma faʻaalia ai faʻasologa o le methylation e le masani ai e fesoʻotaʻi ma tulaga eseese, aemaise lava le kanesa. E ala i le fa'aaogaina o le WGBS, e mafai ai e saienitisi ona maua ni fa'amatalaga e le mafaatusalia i le genome-wide methylation landscapes, ma maua ai se malamalamaga fa'apitoa i le epigenetic mechanisms e fa'avaeina ai le tele o gaioiga fa'aletino ma fa'ama'i.
Vaega Auaunaga
● Manaomia se genome faasinomaga.
● Lambda DNA faʻaopoopo e mataʻituina le lelei o le liua o le bisulfite.
● Fa'asologa ile Illumina NovaSeq.
Tulaga Auaunaga
●Tulaga Auro mo Su'esu'ega Methylation DNA: O lenei tekinolosi faʻaliliuina o le methylation matua e maualuga le saʻo ma toe faʻaleleia lelei.
●I'uga Lautele ma I'uga Fa'avae Tasi:su'esu'eina o nofoaga methylation i le genome-lautele tulaga.
●Fa'atonu atoatoa:tu'uina atu le 'au'aunaga sili ona lelei e tasi mai le fa'ata'ita'iga, fau faletusi, fa'asologa i su'esu'ega bioinformatics.
●Tomai Tele: fa'atasi ai ma galuega fa'asologa o le WGBS ua fa'amae'aina lelei i le tele o ituaiga ituaiga, BMKGENE e maua ai le sili atu ma le sefulu tausaga o le poto masani, o se 'au su'esu'e sili ona tomai, fa'amatalaga atoatoa, ma le lagolago sili ona lelei pe a mae'a fa'atau.
●Avanoa e Auai ma Transcriptomics Analysis: fa'ataga mo le su'esu'ega tu'ufa'atasia o le WGBS ma isi fa'amatalaga omics pei ole RNA-seq.
Fa'ata'ita'iga Fa'amatalaga
| Faletusi | Fuafuaga Fa'asologa | Fautuaina fa'amatalaga fa'amatalaga | Pulea lelei |
| Bisulfite togafitia | Illumina PE150 | 30x loloto | Q30 ≥ 85% Liuga ole bisulfite > 99% |
Fa'ata'ita'iga Manaoga
| Fa'atotonuga (ng/µL) | Aofa'iga (µg) | Manaoga faaopoopo | |
| Genomic DNA | ≥ 5 | ≥ 400 ng | Fa'atapula'aina le fa'aleagaina po'o le fa'aleagaina |
Auala o Galuega Au'aunaga
Fa'ata'ita'iga tu'uina atu
Su'ega DNA
Fausia faletusi
Fa'asologa
Iloiloga fa'amaumauga
Tuuina atu o faʻamatalaga
E aofia ai le au'ili'iliga nei:
● Fa'atonuga lelei ole fa'asologa;
● Faafanua e faasino i le genome;
● Su'esu'ega o faavae methylated 5mC;
● Iloiloga o le tufatufaina o le methylation ma faʻamatalaga;
● Su'esu'ega o Itulagi Eseese Methylated (DMRs);
● Fa'amatalaga aoga o kenera e feso'ota'i ma DMRs.
5mC methylation detection: ituaiga o nofoaga methylated
Methylation faafanua. 5mC methylation genome-lautele tufatufaina
Faʻamatalaga o itulagi maualuga methylated
Itulagi Methylated Eseese: kenera fesootai
Itulagi Methylated Eseese: fa'amatalaga o kenera e feso'ota'i (Gene Ontology)
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.
Fan, Y. et al. (2020) 'Suʻesuʻega o faʻamatalaga DNA methylation i le taimi o le atinaʻeina o maso o le skeletal mamoe e faʻaaoga ai le faʻasologa atoa-genome bisulfite',BMC Genomics, 21(1), itulau 1–15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'O le deoxyribonucleic acid methylation faʻalavelave i tagata faigaluega faʻaalia i le vinyl chloride',Toxicology ma Alamanuia Soifua Maloloina, 38(7), itulau 377–388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Sootaga i le va o le genome methylation, tulaga o RNA e le faʻamaonia, mRNAs ma metabolites i le fua o tamato fua',Le Talaaga o La'au, 103(3), itulau 980–994. doi: 10.1111/TPJ.14778.
