Mbelata Nnọchiteanya Bisulfite Sequencing (RRBS)
Mbelata Nnọchiteanya Bisulfite Sequencing (RRBS) apụtala dị ka ihe dị ọnụ ahịa na-arụ ọrụ nke ọma na usoro dum Genome Bisulfite Sequencing (WGBS) na nyocha DNA methylation. Ọ bụ ezie na WGBS na-enye nghọta zuru oke site n'inyocha genome dum n'otu mkpebi ntọala, ọnụ ahịa ya dị elu nwere ike ịbụ ihe na-egbochi ya. RRBS na-ebelata ihe ịma aka a n'ụzọ dabara adaba site n'ịhọrọ nyocha akụkụ nke genome. Usoro a na-adabere na mmụba nke mpaghara agwaetiti CpG nwere ụbara site na mpịaji MspI sochiri nha nha nke 200-500/600 bps. N'ihi ya, ọ bụ naanị mpaghara dị nso na agwaetiti CpG ka a na-ahazi usoro, ebe ndị nwere agwaetiti CpG dị anya ka ewepụrụ na nyocha ahụ. Usoro a, jikọtara ya na usoro nke bisulfite, na-enye ohere maka nchọpụta dị elu nke DNA methylation, na usoro nhazi usoro, PE150, na-elekwasị anya kpọmkwem na njedebe nke ntinye karịa n'etiti, na-abawanye arụmọrụ nke profaịlụ methylation. RRBS bụ ngwá ọrụ bara uru nke na-enyere nyocha DNA methylation dị ọnụ ahịa aka na ọganihu ihe ọmụma nke usoro epigenetic.
Atụmatụ ọrụ
● Chọrọ mkpụrụ ndụ ihe nketa.
● A na-eji DNA Lambda iji nyochaa arụmọrụ ntụgharị bisulfite.
● A na-enyochakwa arụmọrụ mgbari MspI.
● Mgbari enzyme ugboro abụọ maka ihe atụ osisi.
● Usoro na Illumina NovaSeq.
Uru ọrụ
●Nhọrọ dị ọnụ ahịa na nke ọma na WGBS: na-enyere aka nyocha a ga-eme ya na ọnụ ala dị ala yana ihe achọrọ dị ala.
●Platform zuru ezu:na-enye ọrụ dị mma nke otu nkwụsị site na nhazi nlele, iwu ụlọ akwụkwọ, na usoro na nyocha bioinformatics.
●Ọkachamara sara mbara: na RRBS sequencing oru ngo mezuru nke ọma n'ofe dị iche iche nke ụdị, BMKGENE na-eweta ihe karịrị afọ iri nke ahụmahụ, a ukwuu nkà nyocha otu, keukwu ọdịnaya, na magburu onwe post-ire nkwado.
Nkọwapụta ọrụ
| Ụlọ akwụkwọ | Usoro usoro | Mbupute data akwadoro | njikwa mma |
| MspI gbarie yana ọba akwụkwọ ọgwụgwọ Bisulfite | Ilumina PE150 | 8 gb | Q30 ≥ 85% Ntughari Bisulfite> 99% MspI mbelata arụmọrụ> 95% |
Ihe atụ chọrọ
| Ntinye uche (ng/µL) | Ngụkọta ego (µg) |
| |
| Genomic DNA | ≥ 30 | ≥ 1 | Mmebi ma ọ bụ mmetọ nwere oke |
Usoro ọrụ ọrụ
Nbufe ihe atụ
Ịrụ ụlọ akwụkwọ
Usoro
Nyocha data
Mgbe-ire ọrụ
Ọ gụnyere nyocha ndị a:
● njikwa mma nke usoro ihe ọkụkụ;
● Map na-ezo aka na genome;
● Nchọpụta nke 5mC methylated bases na njirimara motif;
● Nyocha nke nkesa methylation na atụnyere ihe atụ;
● Nyocha nke Mpaghara Methylated dị iche iche (DMR);
● Nkọwa ọrụ nke mkpụrụ ndụ ihe nketa jikọtara na DMR.
Njikwa ogo: arụmọrụ mgbari nri (na maapụ genome)
Njikwa ogo: ntụgharị bisulfite (na mmịpụta ozi methylation)
Map Methylation: 5mC methylation genome-obosara nkesa
Ntụnyere nlele: Nyocha akụrụngwa nke isi
Ntụle mpaghara Methylated dị iche iche (DMRs): map okpomọkụ
Nyochaa ọganihu nyocha nke BMKGene dum genome bisulfite sequencing ọrụ na-akwado site na nchịkọta mbipụta echekwabara.
Li, Z. et al. (2022) 'Mmeghari ntụkwasị obi dị elu n'ime sel ndị dị ka Leydig site na ntinye CRISPR na ihe paracrine',PNAS Nexus, 1 (4). doi: 10.1093/PNASNEXUS/PGAC179.
Tian, H. et al. (2023) 'Nnyocha DNA methylation nke ihe mejupụtara ahụ na ejima monozygotic nke China',Akwụkwọ akụkọ European Journal of Clinical Investigation, 53 (11), p. e14055. doi: 10.1111/ECI.14055.
Wu, Y. et al. (2022) 'DNA methylation na ukwu-na-hip ruru: ọmụmụ ihe gbasara epigenome-wide Association na Chinese monozygotic ejima',Akwụkwọ akụkọ Endocrinological Investigation, 45 (12), p. 2365–2376. doi: 10.1007/S40618-022-01878-4.
