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Hi-C yakavakirwa Chromatin Kudyidzana
Hi-C inzira yakagadzirirwa kubata genomic kumisikidzwa nekubatanidza kuongorora kudhuze-kwakavakirwa kupindirana uye yakakwirira-kuburikidza sequencing. Iyo nzira yakavakirwa pachromatin crosslinking ne formaldehyde, inoteverwa nekugaya uye kubatanidza zvakare nenzira yekuti zvimedu zvakabatana zvakabatana zvinogadzira zvigadzirwa zve ligation. Nekuteedzera izvi zvigadzirwa zve ligation, zvinokwanisika kudzidza iyo 3D sangano reiyo genome. Hi-C inogonesa kudzidza kugoverwa kwezvikamu zvejenome izvo zvakapetwa zvishoma (A compartments, euchromatin) uye zvakanyanya kunge zvichinyorwa, uye matunhu akanyanya kuzara (B compartments, Heterochromatin). Hi-C inogona zvakare kushandiswa kunongedza Topologically Associated Domains (TADs), matunhu ejenome akapeta zvimiro uye anogona kunge aine maitiro ekutaura akafanana, uye kuona chromatin loops, dunhu reDNA rakasungirirwa pamwechete nemapuroteni uye ari kazhinji kupfumiswa muzvinhu zvinodzorwa. BMKGene's Hi-C sequencing sevhisi inopa simba vatsvakurudzi kuti vaongorore zvimiro zvegenomics, vachivhura nzira nyowani dzekunzwisisa genome regulation uye zvazvinoreva muhutano nechirwere.
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Chromatin Immunoprecipitation Sequencing (ChIP-seq)
Chromatin Immunoprecipitation (CHIP) inyanzvi inosimudzira masoja ekudzivirira chirwere kuti apfumese mapuroteni anosunga maDNA uye zvinangwa zvawo zvinoenderana nema genomics. Kubatanidzwa kwayo neNGS kunoita kuti genome-wide profiling yeDNA zvinangwa zvine chekuita nehistone modification, transcription factors, uye mamwe mapuroteni anosunga DNA. Iyi nzira ine simba inogonesa kuenzanisa kwenzvimbo dzinosunga kune akasiyana siyana maseru marudzi, tishu, kana mamiriro. Zvishandiso zveChIP-Seq zvinotora kubva pakudzidza kunyoreswa mutemo uye nzira dzekusimudzira kusvika pakujekesa nzira dzechirwere, zvichiita kuti ive chishandiso chakakosha pakunzwisisa genomic regulation landscapes uye kufambira mberi kweruzivo rwekurapa.
Platform: Illumina NovaSeq
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Whole genome bisulfite sequencing (WGBS)
Whole Genome Bisulfite Sequencing (WGBS) inomira senzira yegoridhe-yakajairwa yekuongorora kwakadzama kweDNA methylation, kunyanya chinzvimbo chechishanu mu cytosine (5-mC), yakakosha inodzora yejini kutaura uye cellular chiitiko. Nheyo iri pasi peWGBS inosanganisira kurapwa kwebisulfite, kukonzeresa shanduko yeunmethylated cytosines kuita uracil (C kusvika kuU), ichisiya methylated cytosines isina kuchinjika. Iyi nzira inopa imwechete-base resolution, ichibvumira vaongorori kuti vaongorore zvizere iyo methylome uye kufumura abnormal methylation mapatani ane chekuita neakasiyana mamiriro, kunyanya cancer. Nekushandisa WGBS, masayendisiti anogona kuwana nzwisiso isingaenzaniswi mune genome-wide methylation landscapes, ichipa kunzwisisa kwakadzama kwemaitiro epigenetic ayo ari pasi pemaitiro akasiyana ebhayoloji nezvirwere.
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Assay yeTransposase-Inowanikwa Chromatin ine Yakakwira Kuburikidza Sequencing (ATAC-seq)
ATAC-seq inzira yepamusoro-yekutevedzana inoshandiswa pakuongorora kuwanikwa kwegenome-wide chromatin. Inoshandisa inopa kunzwisisa kwakadzama kwemaitiro akaomarara epasi rose epigenetic kutonga pamusoro pekutaura kwemajini. Iyo nzira inoshandisa hyperactive Tn5 transposase kune panguva imwe chete chidimbu uye tag yakavhurika chromatin matunhu nekuisa sequencing adapta. Inozotevera PCR kukwidziridzwa kunoguma mukugadzirwa kweraibhurari inoteedzana, iyo inobvumira kuzivikanwa kwakazara kweakavhurika chromatin matunhu pasi peiyo nzvimbo-yenguva mamiriro. ATAC-seq inopa maonero akakwana ezvinowanikwa chromatin landscapes, zvakasiyana nenzira dzinongotarisa pane zvinyorwa zvinosungirira masaiti kana nzvimbo dzakagadziriswa histone. Nekuteedzera aya akavhurika chromatin matunhu, ATAC-seq inoratidza matunhu angangoita anoshanda ekudzora kutevedzana uye angangoita transcription factor inosunga masaiti, ichipa ruzivo rwakakosha mukusimba kwekuchinja kwemajini kutenderera kune genome.
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Yakaderedzwa Representation Bisulfite Sequencing (RRBS)
Yakaderedzwa Representation Bisulfite Sequencing (RRBS) yabuda senzira inodhura uye inoshanda kune Whole Genome Bisulfite Sequencing (WGBS) muDNA methylation research. Nepo WGBS ichipa nzwisiso yakazara nekuongorora iyo genome yese pane imwechete base resolution, mutengo wayo wakakwirisa unogona kuve unomisa chinhu. RRBS zvine hungwaru inodzikamisa dambudziko iri nekusarudza kuongorora chikamu chinomiririra chejenome. Iyi nzira inotsamira pakupfumiswa kweCpG chitsuwa-akapfuma matunhu neMspI cleavage inoteverwa nehukuru kusarudzwa kwe200-500/600 bps zvimedu. Nekuda kweizvozvo, matunhu chete ari pedyo neCpG zvitsuwa anoteedzana, nepo iwo ane zviri kure CpG zvitsuwa asingabatanidzwe kubva mukuongororwa. Iyi nzira, yakasanganiswa ne bisulfite sequencing, inobvumira kuonekwa kwepamusoro-soro yeDNA methylation, uye nzira yekutevedzana, PE150, inotarisa zvakananga kumagumo ekuisa panzvimbo yepakati, kuwedzera kushanda kwemethylation profiling. Iyo RRBS chishandiso chakakosha chinogonesa inodhura-inoshanda DNA methylation tsvakiridzo uye inosimudzira ruzivo rwe epigenetic maitiro.
