Ukuhlolwa kwe-Chromatin efinyeleleka kalula ye-Transposase ene-High Throughput Sequencing (ATAC-seq)
Izici Zesevisi
● Isevisi idinga amasampula ezicubu, esikhundleni sama-nucleic acid akhishwe, ukuze kulondolozwe ukusebenzisana kwe-DNA-protein.
● Indlela ye-ATAC ihlanganisa ukuhlukaniswa kwezicubu, kulandelwe ukuhlukaniswa kwama-nuclei, ukwelashwa nokuhlanzwa kwe-Tn5, ukukhulisa i-PCR, ukukhethwa kosayizi kanye nokulandelana.
● Ukulandelana kwezithombe ku-Illumina NovaSeq
Izinzuzo Zesevisi
●Ukuzwela Okuphezulu:Inani eliphansi lamaseli okuqala lanele ukulungiselela nokulandelanisa umtapo wolwazi.
●Indlela Efundisa Kakhulu: kwembula ngasikhathi sinye izindawo ze-genomic ze-chromatin evulekile kanye nezakhi zokulawula ezisebenzayo njengezindawo zokubopha izinto ezibhaliwe.
●Ukuphindaphinda Okuhle Kokuhlola: ukukopishwa kobuchwepheshe kubonisa ukuphindaphindeka okuhle kakhulu.
●Ubuchwepheshe Obubanzi: Njengoba amakhulu amaphrojekthi okulandelana kwe-ATAC eqediwe ngempumelelo, i-BMKGENE iletha ulwazi olungaphezu kweminyaka eyishumi, ithimba lokuhlaziya elinekhono eliphezulu, okuqukethwe okuphelele, kanye nokusekelwa okuhle kakhulu ngemuva kokuthengisa.
●Kungenzeka Ukujoyina Ukuhlaziywa Kwe-Transcriptomics: okuvumela ukuhlaziywa okuhlanganisiwe kwe-ATAC seq neminye idatha ye-omics njenge-RNA-seq.
● Ukuhlaziywa Okuphelele Kwe-Bioinformatic: okuvumela hhayi kuphela ukuhlonza izindawo ezivulekile ze-chromatin kanye nokusebenza kwazo okuhambisanayo (abagqugquzeli, ama-UT, ama-exons, ama-intron) kodwa futhi nokuhlaziywa komehluko phakathi kwezindawo ezivulekile ze-chromatin phakathi kwamasampula.
Imininingwane Yesevisi
| Umtapo Wolwazi | Isu Lokulandelana | Ukukhishwa kwedatha okunconyiwe | Ikhwalithi yokulawula |
| I-ATAC-seq | I-Illumina PE150 | > 20M okufundwayo Kuye ngobukhulu be-genome (Umuntu: 50 M ufunda) | Q30≥85% |
Izidingo Zesampula
Uhlobo lwamasampula: Izicubu, amangqamuzana aphilayo noma aqandisiwe, igazi
● Inombolo yeselula: ≥ 106amaseli
● Isisindo sezicubu: ≥ 200mg izicubu ezintsha
● Igazi: ≥ 4 mL
Ukugeleza Komsebenzi Wesevisi
Umklamo wokuhlola
Ukulethwa kwesampula
Ukukhishwa kwe-DNA
Ukwakhiwa komtapo wolwazi
Ukulandelana
Ukuhlaziywa kwedatha
Izinsizakalo zangemuva kokuthengisa
Kufaka phakathi Ukuhlaziywa Okulandelayo:
● Ukulawulwa kwekhwalithi yedatha eluhlaza;
● Ukubiza okuphezulu okusekelwe ekudwebeni i-genome yokubhekisela;
● Isichasiselo se-gene endaweni ephakeme;
● Ukuhlaziywa kwe-Motif: ukuhlonza izindawo zokubopha izinto ezibhaliwe (TFBS);
● Ukuhlaziywa Kwezinga Elihlukile kanye nesichasiselo.
1. I-Heatmap ku-ATAC ifunda ukusatshalaliswa ku-TSS nasezifundeni eziseduze (±3 kb)
2. Ukusatshalaliswa kwesifunda se-chromatin esivulekile ezingxenyeni ezahlukene ze-genome
3. Ukwehlukahlukana kokubiza okuphezulu phakathi kwamaqembu
Hlola intuthuko yocwaningo eyenziwe yizinsizakalo zokulandelana kwe-BMKGene ze-ATAC ngokusebenzisa iqoqo lezincwadi ezikhethiwe.
Fu, A.nabanye.(2023) 'Ukuhlanganiswa kwe-genome ye-Telomere-kuya-telomere ye-bitter melon (i-Momordica charantia L. var. abbreviata Ser.) kwembula ukuthuthukiswa kwezithelo, ukwakheka kanye nezici zofuzo zokuvuthwa',Ucwaningo Lwezolimo, 10(1). doi: 10.1093/HR/UHAC228.
UGong, B.nabanye.(2023) 'Ukuvuselelwa kwe-epigenetic kanye nokubhalwa kwe-transcriptional kwe-secretory kinase FAM20C njenge-oncogene ku-glioma',Ijenali Yezakhi Zofuzo Nezakhi Zofuzo, 50(6), amakhasi 422-433. doi: 10.1016/J.JGG.2023.01.008.
Yena, Y.nabanye.(2023) 'I-Butyrate ibuyisela emuva ukumelana kwe-ferroptosis kumdlavuza we-colorectal ngokubangela ukucindezelwa kwe-xCT okuncike ku-c-Fos',I-Redox Biology, 65, ikhasi 102822. doi: 10.1016/J.REDOX.2023.102822.
