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Hi-C tushen hulɗar Chromatin
Hi-C wata hanya ce da aka ƙera don ɗaukar daidaitawar kwayoyin halitta ta hanyar haɗa ma'amala ta tushen kusanci da babban tsarin aiwatarwa. Hanyar ta dogara ne akan chromatin crosslinking tare da formaldehyde, biye da narkewa da sake sakewa ta hanyar da kawai gutsuttsura waɗanda ke da alaƙa da haɗin gwiwa zasu samar da samfuran ligation. Ta hanyar tsara waɗannan samfuran ligation, yana yiwuwa a yi nazarin ƙungiyar 3D na genome. Hi-C yana ba da damar nazarin rarraba sassan kwayoyin halittar da aka cika da sauƙi (A compartments, euchromatin) kuma mafi kusantar yin aiki a rubuce, da kuma yankunan da suka fi cunkoso (B compartments, Heterochromatin). Hakanan ana iya amfani da Hi-C don nunawa Topologically Associated Domains (TADs), yankuna na genome waɗanda suka naɗe da tsarin kuma suna iya samun nau'ikan maganganu iri ɗaya, da kuma gano madaukai na chromatin, yankuna DNA waɗanda sunadaran sunada alaƙa da su kuma waɗanda suke. sau da yawa wadatar da abubuwa masu tsari. Sabis ɗin jerin abubuwan Hi-C na BMKGene yana ba masu bincike damar bincika sararin sararin samaniya na genomics, buɗe sabbin hanyoyi don fahimtar ƙa'idodin kwayoyin halitta da tasirinsa a cikin lafiya da cuta.
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Chromatin Immunoprecipitation Sequencing (ChiP-seq)
Chromatin Immunoprecipitation (CHIP) wata dabara ce da ke ba da damar rigakafin ƙwayoyin cuta don zaɓar sunadaran da ke ɗaure DNA da maƙasudin abubuwan da suka dace. Haɗuwa da shi tare da NGS yana ba da damar fa'idar siginar kwayoyin halittar DNA da ke da alaƙa da gyare-gyaren tarihi, abubuwan rubutu, da sauran sunadaran da ke ɗaure DNA. Wannan ƙwaƙƙwarar hanya tana ba da damar kwatancen wuraren ɗaure a cikin nau'ikan tantanin halitta, kyallen takarda, ko yanayi daban-daban. Aikace-aikacen ChIP-Seq sun bambanta daga nazarin ƙa'idodin rubutu da hanyoyin haɓakawa zuwa haɓaka hanyoyin cututtuka, mai da shi kayan aiki mai mahimmanci don fahimtar yanayin tsarin tsarin kwayoyin halitta da haɓaka hangen nesa na warkewa.
Platform: Illumina NovaSeq
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Gabaɗaya jerin kwayoyin halitta bisulfite (WGBS)
Duka Genome Bisulfite Sequencing (WGBS) yana tsaye a matsayin tsarin daidaitaccen zinari don zurfin bincike na DNA methylation, musamman matsayi na biyar a cikin cytosine (5-mC), mai mahimmanci mai daidaita maganganun kwayoyin halitta da ayyukan salula. Ka'idar da ke ƙarƙashin WGBS ta ƙunshi jiyya na bisulfite, haifar da jujjuyawar cytosines marasa daidaituwa zuwa uracil (C zuwa U), yayin barin cytosin methylated ba canzawa. Wannan dabarar tana ba da ƙuduri mai tushe guda ɗaya, yana bawa masu bincike damar bincikar methylome gabaɗaya kuma su gano tsarin methylation mara kyau da ke da alaƙa da yanayi daban-daban, musamman kansa. Ta hanyar amfani da WGBS, masana kimiyya za su iya samun haske mara misaltuwa cikin yanayin shimfidar wurare masu faɗi na methylation, suna ba da cikakkiyar fahimta game da hanyoyin epigenetic waɗanda ke haifar da matakai da cututtuka iri-iri.
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Ƙididdiga don Canjawa-Sarfafa Chromatin tare da Babban Sabis na Sabis (ATAC-seq)
ATAC-seq babbar dabara ce ta jerin abubuwan da aka yi amfani da ita don nazarin isa ga chromatin mai faɗin kwayoyin halitta. Ana amfani da shi yana ba da zurfin fahimta game da hadaddun hanyoyin sarrafa epigenetic na duniya akan maganganun kwayoyin halitta. Hanyar tana amfani da jujjuyawar Tn5 mai ƙarfi zuwa ga guntu lokaci guda da yiwa buɗaɗɗen yankuna chromatin ta saka adaftan jeri. Sakamakon haɓakawa na PCR na gaba a cikin ƙirƙirar ɗakin karatu mai ɗorewa, wanda ke ba da damar gano cikakkiyar ganewa na yankuna chromatin da ke buɗe ƙarƙashin takamaiman yanayi-lokaci. ATAC-seq yana ba da cikakkiyar ra'ayi na shimfidar wurare na chromatin mai isa, sabanin hanyoyin da ke mayar da hankali kawai kan wuraren ɗaurin rubutu ko takamaiman yankuna da aka gyara na tarihi. Ta hanyar jera waɗannan yankuna na chromatin da ke buɗe, ATAC-seq yana bayyana yankuna mafi yuwuwar samun jerin tsari na aiki da yuwuwar abubuwan da ke ɗaure rubutu, suna ba da fa'ida mai mahimmanci game da haɓakar yanayin yanayin magana a cikin kwayoyin halitta.
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Rage Matsayin Bisulfite Sequencing (RRBS)
Rage Wakilci Bisulfite Sequencing (RRBS) ya fito a matsayin mai inganci mai tsada da ingantaccen madadin Tsarin Tsarin Halittu na Halitta (WGBS) a cikin binciken DNA methylation. Yayin da WGBS ke ba da cikakkun bayanai ta hanyar nazarin dukkan kwayoyin halitta a ƙudurin tushe guda ɗaya, ƙimar sa mai girma na iya zama abin iyakancewa. RRBS da dabara yana rage wannan ƙalubalen ta hanyar zaɓin nazarin sashin wakilci na kwayoyin halitta. Wannan dabarar ta dogara ne akan haɓaka yankuna masu wadatar tsibiri na CpG ta hanyar tsagewar MspI wanda ke biye da girman zaɓi na 200-500/600 bps. Sakamakon haka, kawai yankuna masu kusanci zuwa tsibiran CpG ne aka jera su, yayin da waɗanda ke da tsibiran CpG masu nisa ba a cire su daga binciken. Wannan tsari, haɗe tare da bisulfite sequencing, yana ba da damar gano babban ƙuduri na DNA methylation, da kuma tsarin tsarin, PE150, yana mai da hankali musamman a kan ƙarshen abubuwan da aka saka maimakon tsakiya, yana ƙara haɓakar haɓakar methylation. RRBS kayan aiki ne mai mahimmanci wanda ke ba da damar bincike na DNA methylation mai tsada mai tsada da haɓaka ilimin hanyoyin epigenetic.
