Methylation sequencing from limiting DNA: embryonic, fixed, and microdissected cells

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 po...

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Veröffentlicht in:Methods (San Diego, Calif.) Calif.), 2002-06, Vol.27 (2), p.108-113
Hauptverfasser: Millar, Douglas S., Warnecke, Peter M., Melki, John R., Clark, Susan J.
Format: Artikel
Sprache:eng
Schlagworte:
3T3 Cells
Animals
Bisulfite
Blastocyst
Cells, Cultured
DNA Methylation
Embryo
Laser capture
Mice
Microdissection
Microtomy
Paraffin
Paraffin Embedding
Polymerase Chain Reaction - methods
Sequence Analysis, DNA - methods
Sulfites - metabolism
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Beschreibung
Zusammenfassung: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.
ISSN:1046-2023
1095-9130
DOI:10.1016/S1046-2023(02)00061-0