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Mmekọrịta Chromatin dabere na Hi-C
Hi-C bụ usoro emebere iji weghara nhazi genomic site na ijikọta mmekọrịta dabere na nso nso yana usoro ntinye dị elu. Usoro a dabere na chromatin crosslinking na formaldehyde, na-esote mgbaze na re-ligation n'ụzọ na ọ bụ naanị iberibe ihe jikọrọ ọnụ ga-emepụta ngwaahịa ligation. Site n'usoro ngwaahịa ligation ndị a, ọ ga-ekwe omume ịmụ nhazi 3D nke genome. Hi-C na-enyere aka n'ịmụ nkesa nke akụkụ nke genome nke a na-anakọta nke ọma (A compartments, euchromatin) na o yikarịrị ka ọ ga-arụ ọrụ transcriptionally, na mpaghara ndị a na-akpachi anya (akụkụ B, Heterochromatin). A pụkwara iji Hi-C mee ka ọ pụta ìhè Topologically Associated Domains (TADs), mpaghara nke genome nke nwere ihe owuwu apịaji ma nwee ike inwe usoro okwu yiri ya, na ịchọpụta loops chromatin, mpaghara DNA nke ndị na-edozi jikọtara ọnụ na nke ahụ bụ. na-emekarị ọgaranya na ihe nchịkwa. Ọrụ usoro Hi-C nke BMKGene na-enye ndị na-eme nchọpụta ike inyocha akụkụ mbara igwe nke genomics, na-emepe ụzọ ọhụrụ maka ịghọta ụkpụrụ genome na ihe ọ pụtara na ahụike na ọrịa.
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Usoro mgbochi mgbochi Chromatin (ChiP-seq)
Chromatin Immunoprecipitation (CHIP) bụ usoro na-enye ọgwụ mgbochi iji mee ka protein na-ejikọta DNA na ebumnuche ha kwekọrọ. Njikọ ya na NGS na-enyere aka ịkọwapụta genome-obosara nke ebumnuche DNA metụtara mgbanwe histone, ihe odide ederede, na protein ndị ọzọ na-ejikọta DNA. Ụzọ a siri ike na-enyere aka ntụnyere saịtị njikọ n'ofe ụdị cell, anụ ahụ, ma ọ bụ ọnọdụ dị iche iche. Ngwa ChIP-Seq sitere na ịmụ usoro ederede na ụzọ mmepe ruo n'ịkọwapụta usoro ọrịa, na-eme ka ọ bụrụ ngwa dị mkpa maka ịghọta mpaghara nhazi mkpụrụ ndụ ihe nketa yana ịkwalite nghọta ọgwụgwọ.
Platform: Illumina NovaSeq
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Usoro genome bisulfite dum (WGBS)
Genome Bisulfite Sequencing (WGBS) na-eguzo dị ka usoro ọkọlọtọ ọla edo maka nyocha miri emi nke DNA methylation, kpọmkwem ọnọdụ nke ise na cytosine (5-mC), onye na-achịkwa mkpụrụ ndụ ihe nketa na ọrụ ekwentị. Ụkpụrụ dị n'okpuru WGBS na-agụnye ọgwụgwọ bisulfite, na-eme ka ntụgharị nke cytosin na-adịghị edozi na uracil (C ruo U), ebe ọ na-ahapụ cytosin methylated agbanweghị. Usoro a na-enye mkpebi naanị otu, na-enye ndị nyocha ohere inyocha methylome nke ọma wee kpughee usoro methylation na-adịghị mma metụtara ọnọdụ dị iche iche, ọkachasị kansa. Site n'iji WGBS were, ndị ọkà mmụta sayensị nwere ike nweta nghọta na-enweghị atụ na mbara ala methylation genome, na-enye nghọta dị nro nke usoro epigenetic nke na-ebute usoro na ọrịa dị iche iche.
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Nyocha maka ntụgharị-Chromatin nwere ike ịnweta ya na usoro ntinye dị elu (ATAC-seq)
ATAC-seq bụ usoro usoro ntinye dị elu ejiri maka nyocha nnweta chromatin obosara. Ọ na-eji na-enye nghọta miri emi nke usoro mgbagwoju anya nke njikwa epigenetic zuru ụwa ọnụ gbasara mkpụrụ ndụ ihe nketa. Usoro a na-eji transposase Tn5 hyperactive n'otu oge ma kpado mpaghara chromatin mepere emepe site na itinye ihe nkwụnye usoro. Mgbakwunye PCR na-esote na-arụpụta ịmepụta ọbá akwụkwọ usoro, nke na-enye ohere maka nchọpụta zuru oke nke mpaghara chromatin mepere emepe n'okpuru ọnọdụ oge-oge. ATAC-seq na-enye echiche zuru oke nke odida obodo chromatin nwere ike ịnweta, n'adịghị ka ụzọ na-elekwasị anya naanị na saịtị njide ihe ederede ma ọ bụ mpaghara akụkọ ihe mere eme gbanwere. Site n'ịhazi mpaghara chromatin ndị a mepere emepe, ATAC-seq na-ekpughe mpaghara o yikarịrị ka ọ ga-enwe usoro iwu na-arụ ọrụ yana saịtị nwere ike ijikọ ederede, na-enye nghọta bara uru na mgbanwe mgbanwe mkpụrụ ndụ ihe nketa n'ofe genome.
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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.
