Hoos udhaca metelaada ee loo yaqaan 'Bispulfite'
Jumladaha metelaadda ee hoos u dhaca (RGBS) ayaa u soo ifbaxay wax qiimo-wax ku ool ah oo wax ku ool ah oo kale oo ku saabsan dhammaan marinnada bisulfite oo dhan (WGBs) ee DNA-da makiinadda DNA-da. In kasta oo WGBs ay bixiso aragtiyo dhameystiran iyadoo lagu imtixaamo gebi ahaanba Genome oo dhan ee xalka hal saldhigga ah, qarashka sare ayaa noqon kara qodobka xaddidan. Rrbs si istiraatiiji ah ayaa lagu mititeyaa caqabaddan iyadoo si xulasho ah ay u falanqeyso qayb matalaya oo ka mid ah hiddaha. Qaabkan ayaa ku tiirsan kobcinta kobcinta gobollada qaniga ah ee CPG Jasiiradda by MSPI Floulage oo ay ku xigto cabirka cabirka ee 200-500 / 600/600 jajabyada BPS. Sidaas awgeed, kaliya gobollada ugu dhawaanshaha Islands-yada CPG waa dabacsanaada, halka kuwa leh jasiiradaha CPG fog laga saaray falanqaynta. Hawshan, oo ay weheliso isku xirka bisulfite, waxay u oggolaaneysaa ogaanshaha qaraarka sare ee DNA-da ee DNA-da, iyo habka isku-darka, PE15, waxay si gaar ah diiradda u saareysaa dhamaadka galka halkii dhexda, kordhinta hufnaanta makaanka. RGBS waa aalad muhiim u ah oo awood u siisa cilmi baarista methylation-ka ee waxtarka leh ee DNA-da iyo horumarka aqoonta farsamooyinka apidenotic.
Astaamaha adeegga
● Waxay u baahan tahay tixraac guud.
● Lambda DNA waxaa loo isticmaalaa in lagu kormeero waxtarka badeecada BISUBFIT.
Firfircoonida dheef--dheerowga MSPI ayaa sidoo kale lala soconayaa.
● Labajibbiska enzyme-ka labajibbiska ah ee tusaalooyinka dhirta.
● Isku xigxiga ifafaalaha Noveviiq.
Faa'iidooyinka adeegga
●Beddelka wax ku oolka ah ee wax ku oolka ah oo hufan ee WGBs: Awood u siinta falanqaynta lagu sameeyo qiimaha hoose iyo shuruudaha muunad hoose.
●Madal buuxa:Sii adeeg aad u wanaagsan oo heer sare ah ka baaraandegida muunadda, dhismaha maktabadda, iyo isku xirka raadinta falanqaynta bioinformormforms.
●Khibrad ballaaran: Mashaariicda isku xidhka rrbs ee si guul leh lagu dhammeeyay noocyo kaladuwan oo kaladuwan, BMKGE waxay keentaa toban sano oo waayo-aragnimo ah, oo ah koox xirfad sare leh, oo ah koox dhammaystiran, iyo taageerada iibka ee ugu fiican.
Faahfaahinta adeegga
| Maktabad | Istaraatiijiyadda isku xigta | Soo saarista xogta lagu taliyay | Xakamaynta Tayada |
| Maktabadda MSPI ee dheefshiidda MSPI iyo maktabadda ee Bisulfite | Iftiimina PE150 | 8 GB | Q30 ≥ 85% Beddelka BISULFIT> 99% MSPI goynta taynta> 95% |
Shuruudaha muunadda
| U fiirsashada (NG / μL) | Wadarta lacagta (μg) |
| |
| DNA Genomic | ≥ 30 | 1 | Nabaad-guur xadidan ama faddarayn |
Socodka shaqada adeegga
Keenista muunad
Dhismaha Maktabadda
Kala xidowdo
Falanqaynta Xogta
Adeegyada ka dib
Waxaa ku jira falanqaynta soo socota:
● Xakamaynta cayriin ee contrient;
● Khariidadda si loo tixraaco tixraaca hiddaha;
● Aasaaska 5mc ee saldhigyada methylated iyo aqoonsiga motaif;
Fist Falanqaynta qaybinta methlation-ka iyo isbarbardhiga muunad;
Falanqaynta Gobollada si kala sooc ah oo methylated (DMRs);
● Qodobka hidda-wadaha ee hawl-galka ah ee la xiriira DMRs.
Xakamaynta Tayada: Kartida dheef-shiidka (ee khariidaynta Genome)
Xakamaynta Tayada: beddelka bisulfite (ee ku dhaca soo-saarista macluumaadka methlation)
Khariidadda methylation: 5mc methylation-ka oo loo qaybiyo
Isbarbardhiga muunad: falanqaynta guud ee guud
Falanqaynta kala duwan ee metrated (DMRS) Falanqaynta: HeatMap
Sahaminta sahaminta cilmi baarista ee ay fududeeyaan adeegyada 'BMKGene's oo dhan ee shirkadaha' Bisulfite 'ee loo yaqaan' Bisulfite 'ee loo yaqaan' Bisulfite 'ee loo yaqaan' Bisulfite 'oo loo maro aruurinta daawooyinka daabacan ee daabacaadaha.
Li, z. et al. (2022) 'daacadnimo sare ayaa canaananaya unugyada Lyadig oo u eg Leydig-sida ay u leeyihiin dhaqdhaqaaqyo downfr ah iyo arrimo kacsan',Pnas nexus, 1 (4). doi: 10.1093 / pnasnexus / pgac179.
Tian, h. et al. (2023) 'Falanqaynta Graad ee DNA-ga oo dhan DNA-ga oo dhan ee kabuujinta jirka ee mataanaha Modozygotic ee Shiinaha',Joornaalka Yurub ee baaritaanka caafimaad ee joogtada ah, 53 (11), p. e14055. doi: 10.1111 / ECI.14055.
WU, Y. Et al. (2022) 'DNA-da' methylation iyo cabir-sip-ka-sip-sip-ka: Daraasad urur oo Epidenome-oo ballaaran oo ku saabsan mataanaha Modozygotic-ka Shiinaha ',Joornaalka baaritaanka endocrinological, 45 (12), pp. 2365-2376. doi: 10.1007 / s40618-022-0187-4.
