Whole genome bisulfite sequencing (WGBS)
Whole Genome Bisulfite Sequencing (WGBS) imayima ngati njira yodziwika bwino ya golide yowunikira mozama DNA methylation, makamaka malo achisanu mu cytosine (5-mC), wowongolera kwambiri wamafotokozedwe amtundu ndi zochitika zama cell. Mfundo yomwe ili pansi pa WGBS imaphatikizapo chithandizo cha bisulfite, kupangitsa kutembenuka kwa ma cytosine osatulutsidwa kukhala uracil (C mpaka U), ndikusiya ma methylated cytosine osasinthika. Njirayi imapereka njira imodzi yokha, yomwe imalola ofufuza kuti afufuze mozama za methylome ndikupeza njira zachilendo za methylation zomwe zimagwirizanitsidwa ndi mikhalidwe yosiyanasiyana, makamaka khansa. Pogwiritsa ntchito WGBS, asayansi atha kupeza zidziwitso zosayerekezeka za mawonekedwe a genome-wide methylation, ndikupereka kumvetsetsa kwapang'onopang'ono kwa njira za epigenetic zomwe zimayambitsa njira zosiyanasiyana zamoyo ndi matenda.
Mawonekedwe a Utumiki
● Pamafunika ma genome.
● Lambda DNA imawonjezedwa kuti iwonetsetse kusinthika kwa bisulfite.
● Kutsata pa Illumina NovaSeq.
Ubwino wa Utumiki
●Muyeso wa Golide wa DNA Methylation Research: Ukadaulo wokhwima wa kutembenuka kwa methylation uli ndi kulondola kwambiri komanso kuberekana kwabwino.
●Kufalikira Kwakukulu ndi Kukhazikika Kwamodzi:kuzindikira kwa malo a methylation pamlingo wa genome-wide.
●Pulatifomu Yathunthu:perekani ntchito imodzi yabwino kwambiri kuyambira pakukonza zitsanzo, kumanga laibulale, kutsata kusanthula kwa bioinformatics.
●Katswiri Wazambiri: ndi ntchito zotsatizana za WGBS zomalizidwa bwino pamitundu yosiyanasiyana ya zamoyo, BMKGENE imabweretsa zokumana nazo zopitilira khumi, gulu laukadaulo laukadaulo, zokhutira, komanso chithandizo chabwino kwambiri cha pambuyo pogulitsa.
●Kuthekera Kujowina ndi Transcriptomics Analysis: kulola kusanthula kophatikizika kwa WGBS ndi data ina ya omics monga RNA-seq.
Zitsanzo Zofotokozera
| Library | Njira Yotsatirira | Kutulutsa kovomerezeka kwa data | Kuwongolera khalidwe |
| Mankhwala a Bisulfite | Chithunzi cha PE150 | 30x kuya | Q30 ≥ 85% Kutembenuka kwa Bisulfite> 99% |
Zitsanzo Zofunika
| Kukhazikika (ng/µL) | Ndalama zonse (µg) | Zofunikira zowonjezera | |
| Genomic DNA | ≥ 5 | ≥ 400 ng | Kuwonongeka pang'ono kapena kuipitsidwa |
Kuyenda kwa Ntchito Yantchito
Kupereka zitsanzo
Kutulutsa kwa DNA
Kumanga laibulale
Kutsata
Kusanthula deta
Kutumiza kwa data
Mulinso zowunikira izi:
● Yaiwisi kandalama kuwongolera khalidwe;
● Kupanga mapu osonyeza ma genome;
● Kuzindikira maziko a 5mC methylated;
● Kuwunika kwa kugawa kwa methylation ndi ndemanga;
● Kusanthula kwa Madera Osiyanasiyana a Methylated (DMRs);
● Ndemanga zogwirira ntchito za majini okhudzana ndi ma DMR.
Kuzindikira kwa 5mC methylation: mitundu yamasamba a methylated
Mapu a Methylation. 5mC methylation genome-wide kugawa
Kufotokozera kwa madera okhala ndi methylated kwambiri
Madera Osiyanasiyana a Methylated: majini ogwirizana
Madera Osiyanasiyana a Methylated: ndemanga zamajini ogwirizana (Gene Ontology)
Onani kupita patsogolo kwa kafukufuku komwe kumayendetsedwa ndi BMKGene's genome bisulfite sequencing services kudzera mgulu lazofalitsa.
Fan, Y. et al. (2020) 'Kusanthula kwa mbiri ya DNA methylation pakukula kwa minofu ya nkhosa pogwiritsa ntchito ma genome bisulfite sequencing' (2020)BMC Genomics, 21(1), tsamba 1-15. doi: 10.1186/S12864-020-6751-5.
Zhao, X. et al. (2022) 'Novel deoxyribonucleic acid methylation zosokoneza mwa ogwira ntchito omwe ali ndi vinyl chloride',Toxicology ndi Industrial Health, 38(7), masamba 377-388. doi: 10.1177/07482337221098600
Zuo, J. et al. (2020) 'Ubale pakati pa genome methylation, milingo ya ma RNA osalemba ma code, mRNAs ndi metabolites pakucha zipatso za phwetekere' (2020)The Plant Journal, 103(3), tsamba 980-994. doi: 10.1111/TPJ.14778.
