Hi-C fa'avae Chromatin Interaction
Vaega Auaunaga
● Fa'asologa ile Illumina NovaSeq ile PE150.
● E mana'omia e le au'aunaga ni fa'ata'ita'iga a le tino, nai lo nucleic acids ua maua mai, e fa'afeso'ota'i ma formaldehyde ma fa'asao ai feso'ota'iga DNA-protein.
● O le suʻesuʻega Hi-C e aofia ai le faʻatapulaʻaina ma le toe faʻaleleia o pito pipii i le biotin, sosoo ai ma le faʻataʻamilomilo o faʻaiʻuga faʻamaʻi aʻo faʻasaoina fegalegaleaiga. Ona toso ifo lea o le DNA i lopa streptavidin ma fa'amamāina mo le tapenaga a le faletusi.
Tulaga Auaunaga
●Fuafuaga Enzyme Fa'atapula'a Sili: ia faʻamautinoa le maualuga o le maualuga o le Hi-C i luga o ituaiga eseese e oʻo atu i le 93% taʻitoʻatasiga aoga.
●Tomai Tele ma Fa'amaumauga Lomiga:BMKGene e tele le poto masani ma> 2000 Hi-C sequencing poloketi mai 800 ituaiga eseese ma pateni eseese. E silia ma le 100 mataupu fa'asalalau fa'atasi ai ma le fa'atuputeleina o a'afiaga e sili atu i le 900.
●Vaega o Bioinformatics sili ona tomai:fa'atasi ai ma pateni i totonu o le fale ma puletaofia polokalame mo su'esu'ega a le Hi-C ma su'esu'ega fa'amaumauga ma se polokalama fa'akomepiuta fa'amatalaga fa'aaliga na faia e le tagata lava ia.
●Lagolago ina ua uma ona faatau atu:O la matou tautinoga e faʻalautele atu i tua atu o le faʻamaeʻaina o galuega faʻatasi ma le 3-masina pe a maeʻa le faʻatau atu vaitaimi auaunaga. I le taimi lea, matou te ofoina atu le tulitatao o poloketi, fesoasoani faʻafitauli, ma Q&A sauniga e faʻafeiloaʻi ai soʻo se fesili e fesoʻotaʻi ma iʻuga.
●Fa'amatalaga Atoa: matou te fa'aogaina le tele o fa'amaumauga e fa'ailoa ai genes ma suiga fa'ailoa ma fa'atino le su'esu'ega fa'atamaoaigaina tutusa, maua ai fa'amatalaga i le tele o galuega su'esu'e.
Fa'amatalaga Auaunaga
| Faletusi | Fuafuaga Fa'asologa | Fautuaina fa'amatalaga fa'amatalaga | I'uga o Fa'ailoga Hi-C |
| Faletusi Hi-C | Illumina PE150 | Chromatin matasele: 150x TAD: 50x | Chromatin Loop: 10Kb TAD: 40Kb |
Manaoga Auaunaga
| Fa'ata'ita'iga ituaiga | Aofa'i mana'omia |
| Aano o manu | ≥2g |
| Toto Atoa | ≥2mL |
| Sigi | ≥1g |
| La'au- tama'i tino | 1g/aliquot, 2-4 aliquotes fautuaina |
| Selula Aganuu | ≥1x107 |
E aofia ai le Su'esu'ega o lo'o mulimuli mai:
● Fa'amatalaga mata'utia QC;
● Fa'afanua ma le faletusi Hi-C QC: ta'ito'alua fegalegaleai fa'amaonia ma Interaction Deay Exponents (IDEs);
● Genome-wide Interaction Profiling: su'esu'ega cis/trans ma fa'afanua feso'ota'iga Hi-C;
● Iloiloga o le tufaina o vaega A/B;
● Fa'ailoaina o TADs ma chromatin loops;
● Su'esu'ega 'ese'ese i elemene fausaga chromatin 3D i fa'ata'ita'iga ma fa'amatalaga fa'atino tutusa o kenera fa'atasi.
Cis ma felauaiga fa'asoa fa'asoa
Fa'afanua vevela o feso'ota'iga chromosomal i le va o fa'ata'ita'iga
Genome-lautele tufatufaina o vaega A/B
Genome-lautele tufatufaina o chromatin matasele
Va'aiga o TADs
Su'esu'e le alualu i luma o su'esu'ega na fa'afaigofieina e le BMKGene's Hi-C sequencing services e ala i le aoina o fa'asalalauga.
Meng, T. et al. (2021) 'O se fa'atusatusaga tu'ufa'atasiga su'esu'ega multi-omics e iloa ai le CA2 o se fa'amoemoega fou mo chordoma',Neuro-Oncology, 23(10), itulau 1709–1722. doi: 10.1093/NEUONC/NOAB156.
Xu, L. et al. (2021) '3D disorganization ma le toe faʻatulagaina o genome e maua ai faʻamatalaga i le pathogenesis o le NAFLD e ala i le tuʻufaʻatasia o le Hi-C, Nanopore, ma le RNA sequencing',Acta Pharmaceutica Sinica B, 11(10), itulau 3150–3164. doi: 10.1016/J.APSB.2021.03.022.
