High sensitivity mapping of methylated cytosines (original) (raw)

Journal Article

,

1

Kanematsu Laboratories, Royal Prince Alfred Hospital

Missenden Road, Camperdown, NSW 2050

2

CSIRO Division of Biomolecular Engineering, Sydney Laboratory

PO Box 184, North Ryde, NSW 2113

*To whom correspondence should be addressed at: CSIRO Division of Biomolecular Engineering, Sydney Laboratory, PO Box 184, North Ryde, NSW 2113, Australia

Search for other works by this author on:

,

1

Kanematsu Laboratories, Royal Prince Alfred Hospital

Missenden Road, Camperdown, NSW 2050

Search for other works by this author on:

,

1

Kanematsu Laboratories, Royal Prince Alfred Hospital

Missenden Road, Camperdown, NSW 2050

Search for other works by this author on:

3

School of Biological Sciences, A12, University of Sydney

NSW 2006, Australia

Search for other works by this author on:

Revision received:

01 July 1994

Published:

11 August 1994

• PDF
• Split View
• • Article contents
  • Figures & tables
  • Video
  • Audio
  • Supplementary Data
• Cite

Cite

J.CIark Susan, Janet Harrison, Cheryl L. Paul, Marianne Frommer, High sensitivity mapping of methylated cytosines, Nucleic Acids Research, Volume 22, Issue 15, 11 August 1994, Pages 2990–2997, https://doi.org/10.1093/nar/22.15.2990
Close

Navbar Search Filter Mobile Enter search term Search

Abstract

An understanding of DNA methylation and its potential role in gene control during development, aging and cancer has been hampered by a lack of sensitive methods which can resolve exact methylation patterns from only small quantities of DNA. We have now developed a genomic sequencing technique which is capable of detecting every methylated cytosine on both strands of any target sequence, using DNA isolated from fewer than 100 cells. In this method, sodium bisulphite is used to convert cytosine residues to uracil residues in single-stranded DNA, under conditions whereby 5-methylcytosine remains non-reactive. The converted DNA is amplified with specific primers and sequenced. All the cytosine residues remaining in the sequence represent previously methylated cytosines in the genome. The work described has defined procedures that maximise the efficiency of denaturetion, bisulphite conversion and amplification, to permit methylation mapping of single genes from small amounts of genomic DNA, readily available from germ cells and early developmental stages.

This content is only available as a PDF.

© 1994 Oxford University Press

I agree to the terms and conditions. You must accept the terms and conditions.

Submit a comment

Name

Affiliations

Comment title

Comment

You have entered an invalid code

Thank you for submitting a comment on this article. Your comment will be reviewed and published at the journal's discretion. Please check for further notifications by email.

Citations

Views

Altmetric

Metrics

Total Views 1,076

238 Pageviews

838 PDF Downloads

Since 11/1/2016

×

Email alerts

Citing articles via

• Latest

• Most Read

• Most Cited

More from Oxford Academic