Kusanthula kwa bisulfite yonse ya genome (WGBS)
Njira imeneyi, yochokera ku bisulfite treatment ya DNA kuti ipangitse kusintha kwa ma cytosines osapangidwa ndi methyl kukhala uracil (C kupita ku U), pomwe kusiya ma cytosines opangidwa ndi methyl osasinthika, ndiyo njira yabwino kwambiri yofufuzira mozama za methylation ya DNA, makamaka malo achisanu mu cytosine (5-mC), chowongolera chofunikira cha kufotokozera majini ndi ntchito zamaselo.
Njirayi imapereka yankho la maziko amodzi, zomwe zimathandiza ofufuza kufufuza mokwanira za methylome ndikupeza njira zachilendo za methylation mkati mwa zitsanzo. Pogwiritsa ntchito WGBS, asayansi amatha kupeza chidziwitso chosayerekezeka pa malo a methylation mu genome yonse, zomwe zimapangitsa kuti amvetsetse bwino njira za epigenetic zomwe zimayambitsa njira zosiyanasiyana zamoyo ndi matenda.
Zinthu Zofunika pa Utumiki
● Kutsatana kwa Illumina NovaSeq.
● Imafuna genome yodziwikiratu.
● DNA ya Lambda yawonjezeredwa kuti iwonetse momwe bisulfite imagwirira ntchito.
Ubwino wa Utumiki
●Muyezo wa Golide wa Kafukufuku wa Methylation wa DNA: Ukadaulo wokhwimawu uli ndi kulondola kwakukulu komanso kubwerezabwereza bwino.
●Kufalikira Kwambiri ndi Kusasinthika kwa Chimodzi:Kuzindikira malo a methylation kumachitika pamlingo wa genome yonse.
●Nsanja Yonse:Timapereka ntchito yokhazikika kuyambira kukonza zitsanzo, kumanga laibulale, kutsata mpaka kusanthula kwa bioinformatics.
●Ukatswiri Wochuluka: Tili ndi mapulojekiti osiyanasiyana omwe adamalizidwa m'mitundu yosiyanasiyana ya zamoyo. BMKGENE imabweretsa chidziwitso cha zaka zoposa khumi, gulu lofufuza laluso kwambiri, zomwe zili mkati, komanso chithandizo chabwino kwambiri pambuyo pogulitsa.
●Kuthekera Kogwirizana ndi Kusanthula kwa Transcriptomics: Kulola kusanthula kophatikizana kwa WGBS ndi deta ina ya omics monga RNA-seq.
Zitsanzo Zofotokozera
| Laibulale | Njira Yotsatizana | Zotsatira zodziwika bwino za deta | Kuwongolera khalidwe |
| Bisulfite yothandizidwa | Illumina PE150 | Kuzama kwa 30x | Q30 ≥ 85% Kusintha kwa Bisulfite > 99% |
Zofunikira pa Zitsanzo
| Kuchuluka kwa madzi (ng/µL) | Chiwerengero chonse (µg) | Zofunikira zina | |
| DNA ya majini | ≥ 5 | ≥ 400 ng | Kuwonongeka kochepa kapena kuipitsidwa |
Kayendedwe ka Ntchito Yotumikira
Kutumiza zitsanzo
Kuchotsa DNA
Kumanga laibulale
Kutsatana
Kusanthula deta
Kutumiza deta
Utumikiwu umaphatikizapo kusanthula kotsatira:
● Kuwongolera khalidwe la kutsata kosaphika;
● Kujambula mapu a majini ofunikira;
● Kuzindikira maziko a methylated a 5mC;
● Kusanthula kwa kugawa kwa methylation ndi kufotokozera;
● Kusanthula kwa Madera Osiyanasiyana a Methylated (DMRs);
● Kufotokozera ntchito ya majini okhudzana ndi ma DMR.
Kuzindikira kwa methylation ya 5mC: mitundu ya malo okhala ndi methylated
Mapu a Methylation. Kugawa kwa 5mC methylation genome lonse
Chidziwitso cha madera omwe ali ndi methyl yambiri
Madera Osiyanasiyana a Methylated: majini ogwirizana nawo
Madera Osiyanasiyana a Methylated: kufotokozera majini ogwirizana nawo (Gene Ontology)
Fufuzani kupita patsogolo kwa kafukufuku komwe kwathandizidwa ndi ntchito zonse za BMKGene zowerengera majini a bisulfite kudzera mu zofalitsa zosiyanasiyana.
Fan, Y. et al. (2020) 'Kusanthula kwa ma DNA methylation profiles panthawi ya kukula kwa minofu ya mafupa a nkhosa pogwiritsa ntchito njira yonse ya bisulfite',BMC Genomics, 21(1), tsamba 1-15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Kusokonezeka kwatsopano kwa deoxyribonucleic acid methylation mwa ogwira ntchito omwe ali ndi vinyl chloride',Kuopsa kwa Zaumoyo ndi Zaulimi, 38(7), masamba 377-388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Ubale pakati pa methylation ya genome, kuchuluka kwa ma RNA osalemba ma code, mRNA ndi metabolites mu zipatso za phwetekere zomwe zikukhwima',Nyuzipepala ya Zomera, 103(3), tsamba 980-994. doi: 10.1111/TPJ.14778.
