“Whole genome bisulfite sequencing”的意思、由来-开放百科全书

Whole genome bisulfite sequencing (WGBS), is a next-generation sequencing technology used to determine the DNA methylation status of single cytosines by treating the DNA with sodium bisulfite before sequencing. Sodium bisulfite is a chemical compound that converts unmethylated cytosines into uracil.^[1]^[2] The cytosines that haven't converted in uracil are methylated. After sequencing, the unmethylated cytosines appear as thymines.

This technique measures single-cytosine methylation levels genome-wide and directly estimates the ratio of molecules methylated rather than enrichment levels. However, this method requires essentially resequencing the entire genome multiple times for every experiment.^[3]

References

1. ^{{Cite journal|last=Frommer|first=M.|last2=McDonald|first2=L. E.|last3=Millar|first3=D. S.|last4=Collis|first4=C. M.|last5=Watt|first5=F.|last6=Grigg|first6=G. W.|last7=Molloy|first7=P. L.|last8=Paul|first8=C. L.|date=1992-03-01|title=A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands|journal=Proceedings of the National Academy of Sciences of the United States of America|volume=89|issue=5|pages=1827–1831|issn=0027-8424|pmc=48546|pmid=1542678|doi=10.1073/pnas.89.5.1827}}
2. ^{{Cite journal|last=Clark|first=S J|last2=Harrison|first2=J|last3=Paul|first3=C L|last4=Frommer|first4=M|date=1994-08-11|title=High sensitivity mapping of methylated cytosines.|journal=Nucleic Acids Research|volume=22|issue=15|pages=2990–2997|issn=0305-1048|pmc=310266|pmid=8065911|doi=10.1093/nar/22.15.2990}}
3. ^{{Cite journal|last=Stevens|first=Michael|last2=Cheng|first2=Jeffrey B.|last3=Li|first3=Daofeng|last4=Xie|first4=Mingchao|last5=Hong|first5=Chibo|last6=Maire|first6=Cécile L.|last7=Ligon|first7=Keith L.|last8=Hirst|first8=Martin|last9=Marra|first9=Marco A.|date=2013-09-01|title=Estimating absolute methylation levels at single-CpG resolution from methylation enrichment and restriction enzyme sequencing methods|journal=Genome Research|volume=23|issue=9|pages=1541–1553|doi=10.1101/gr.152231.112|issn=1088-9051|pmc=3759729|pmid=23804401}}