Profiling DNA methylation by bisulfite genomic sequencing: problems and solutions.
The surge of interest in DNA methylation during the last two decades has triggered an urgent need for an effective method to detect the methylation status of the cytosines in the genome. Bisulfite genomic sequencing is the most attractive choice so far for many laboratories. Various protocols have been established, but difficulties are often encountered, particularly by individuals who have limited experience in this field. This analysis presents a simple protocol that has consistently worked well in our laboratory. Discussions of potential technical problems and corresponding solutions are also included to facilitate the reproducibility of this protocol.
Department of Biology, University of Alabama at Birmingham, USA.
Liang
L
Liu
Rebecca C
RC
Wylie
Nathaniel J
NJ
Hansen
Lucy G
LG
Andrews
Trygve O
TO
Tollefsbol
D011232:Chemical Precipitation; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D019175:DNA Methylation; D023281:Genomics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000737:chemistry*
D013447:Sulfites; D004247:DNA