Go Back   SEQanswers > Literature Watch

Similar Threads
Thread Thread Starter Forum Replies Last Post
PubMed: Sequential DNA methylation changes are associated with DNMT3B overexpression Newsbot! Literature Watch 0 11-12-2010 03:00 AM
PubMed: Relationship between nucleosome positioning and DNA methylation. Newsbot! Literature Watch 0 06-01-2010 03:00 AM
PubMed: Analysing diet of small herbivores: the efficiency of DNA barcoding coupled w Newsbot! Literature Watch 0 08-22-2009 06:11 AM
PubMed: Profiling DNA methylation from small amounts of genomic DNA starting material Newsbot! Literature Watch 0 11-07-2008 10:30 AM

Thread Tools
Old 09-14-2011, 03:00 AM   #1
RSS Posting Maniac

Join Date: Feb 2008
Posts: 1,443
Default PubMed: Analysing DNA methylation using bisulphite pyrosequencing.

Syndicated from PubMed RSS Feeds

Analysing DNA methylation using bisulphite pyrosequencing.

Methods Mol Biol. 2011;791:33-53

Authors: Mikeska T, Felsberg J, Hewitt CA, Dobrovic A

Bisulphite pyrosequencing is a quantitative methodology for the investigation of DNA methylation of sequences up to 100-bp in length. Biotin-labelled, single-stranded PCR products generated from bisulphite-treated DNA are used as a template with an internal primer to perform the pyrosequencing reaction. Nucleotides are added in a predetermined order in each pyrosequencing cycle and the amount of incorporated nucleotide results in a proportional emission of light. DNA methylation ratios are calculated from the levels of light emitted from each nucleotide incorporated at individual CpG positions in a strand-dependent manner. The methylation detection limit at individual CpG sites is approximately 5% and the results are displayed as an average methylation level for each CpG position assayed across all amplification products generated during a PCR reaction. As a consequence, bisulphite pyrosequencing allows the identification of heterogeneous DNA methylation patterns but does not provide information at a single allele resolution. This methodology is suited to analyse short DNA sequences such as those typically extracted from formalin-fixed paraffin-embedded specimens. Nevertheless, longer PCR products can be sequenced by serial bisulphite pyrosequencing, which utilises tandem assays along the amplicon. The general information provided is applicable for all formats of current pyrosequencing instruments, however, a specific protocol for the PyroMark Q24 instrument is provided.

PMID: 21913070 [PubMed - in process]

Newsbot! is offline   Reply With Quote

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off

All times are GMT -8. The time now is 09:16 AM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2021, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO