Accurate sodium bisulfite sequencing in plants.
DNA cytosine methylation is a conserved epigenetic modification frequently correlating with transcriptional silencing in a wide variety of eukaryotic organisms. Sodium bisulfite treatment of DNA converts unmethylated cytosine to uracil, while 5-methylated cytosine is protected. We describe techniques that ensure reliable sequencing data following sodium bisulfite conversion and to avoid common pitfalls such as amplification of unconverted DNA and inclusion of sibling clones.
Henderson
Ian R
IR
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge, UK.
Chan
Simon R
SR
Cao
Xiaofeng
X
Johnson
Lianna
L
Jacobsen
Steven E
SE
D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D003596:Cytosine / Q000737:chemistry; D019175:DNA Methylation / Q000235:genetics; D044127:Epigenesis, Genetic; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D013447:Sulfites / Q000494:pharmacology*; D014498:Uracil / Q000737:chemistry
D013447:Sulfites; D014498:Uracil; D003596:Cytosine; C009279:sodium bisulfite