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DNA methylation levels in Drosophila melanogaster are nearly undetectable. [86] ... which is a high-throughput genome-wide analysis of DNA methylation.
The first few steps of COBRA, and the molecular changes caused by each step to methylated and unmethylated CpG sites. Combined Bisulfite Restriction Analysis (or COBRA) is a molecular biology technique that allows for the sensitive quantification of DNA methylation levels at a specific genomic locus on a DNA sequence in a small sample of genomic DNA. [1]
It was recently shown that cytosine methylation patterns tend to be concordant over short (~1 kb) regions. [2] The patterns represented by the HpaII sites therefore tend to be representative of other CG dinucleotides locally. The analysis of HELP data involves quality analysis and normalization.
All subsequent DNA methylation analysis techniques using bisulfite-treated DNA is based on this report by Frommer et al. (Figure 2). [6] Although most other modalities are not true sequencing-based techniques, the term "bisulfite sequencing" is often used to describe bisulfite-conversion DNA methylation analysis techniques in general.
Reduced representation bisulfite sequencing (RRBS) is an efficient and high-throughput technique for analyzing the genome-wide methylation profiles on a single nucleotide level. It combines restriction enzymes and bisulfite sequencing to enrich for areas of the genome with a high CpG content.
The Illumina Methylation Assay using the Infinium I platform uses 'BeadChip' technology [clarification needed] to generate a comprehensive genome-wide profiling of human DNA methylation. Similar to bisulfite sequencing and pyrosequencing , this method quantifies methylation levels at various loci within the genome .
The mammalian genome naturally contains DNA methylation, but only at CpG sites, so GpC methylation can be differentiated from genomic methylation after bisulfite sequencing. This allows simultaneous analysis of the nucleosome footprint and endogenous methylation on the same DNA molecules. [1]
The HRM methodology has also been exploited to provide a reliable analysis of the methylation status of DNA. This is of significance since changes to the methylation status of tumour suppressor genes, genes that regulate apoptosis and DNA repair, are characteristics of cancers and also have implications for responses to chemotherapy.