Methylation sequencing from limiting DNA: embryonic, fixed, and microdissected cells.
It is frequently useful to determine the methylation state of samples containing limited amounts of DNA such as from embryos, or from fixed tissue samples in which DNA is degraded or difficult to isolate. By modification of the standard protocols for DNA preparation and bisulfite treatment, it is possible to obtain DNA methylation sequence data for such samples. We present methods for bisulfite treatment of embryos, fixed sections, and samples obtained by laser capture microdissection, and discuss the additional experimental considerations required when working with small numbers of cells or degraded DNA samples.
Millar
Douglas S
DS
Kanematsu Laboratories, Sydney Cancer Centre, Royal Prince Alfred Hospital, Missenden Road, Camperdown, New South Wales 2050, Australia.
Warnecke
Peter M
PM
Melki
John R
JR
Clark
Susan J
SJ
D016475:3T3 Cells; D000818:Animals; D001755:Blastocyst; D002478:Cells, Cultured; D019175:DNA Methylation; D051379:Mice; D008867:Microtomy; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000378:metabolism
D013447:Sulfites