MethylQuant is a cost-effective and relatively simple technique which enables quantitative analysis of the methylation status of a single cytosine at specific positions in DNA that can be assimilated to the quantitative detection of a single nucleotide polymorphism (SNP). After bi ...
DNA methylation in mammals has been shown to play many important roles in diverse biological phenomena. Here we describe a simple and straightforward method that quantitatively measures site-specific levels of DNA methylation in a quick and cost-effective manner. The quantitative a ...
Two approaches for the evaluation of the relative degree of global DNA methylation through the quantification of 2′ deoxynucleosides are described. Detection and quantification of 5-methyl 2′-deoxycytidine in genomic DNA is performed using both high-performance capillary ele ...
DNA methylation is the best-studied epigenetic modification, and in mammals it describes the conversion of cytosine to 5-methylcytosine in the context of CpG dinucleotides. In recent years, it has become evident that epigenetic mechanisms are severely disrupted in human neoplasia, a ...
As the role for epigenetic signals in genome regulation becomes increasingly understood, the ability to accurately measure levels of DNA methylation at individual cytosines throughout the genome is becoming increasingly important. In contrast to traditional methods for the qua ...
The analysis of epigenetic changes in genomic DNA has seen an exponentially increasing interest over the last years. Within the field of epigenetics DNA methylation patterns have become of particular interest to the scientific community. The covalent addition of a methyl group to cytosi ...
Most available protocols for gene-specific DNA methylation analysis are either labor intensive, not quantitative, or limited to the measurement of the methylation status of only one or very few CpG positions. Pyrosequencing is a real-time sequencing technology that overcomes these ...
DNA methylation is an essential epigenetic modification in the human genome. For the investigation of DNA methylation patterns, bisulfite conversion and DNA sequencing is a method of choice, because it provides detailed information on the methylation pattern of individual DNA mole ...
Epigenetic silencing of a gene can be reversed, resulting in reactivation of expression, by drugs such as the DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5Aza-dC, azacytidine). This drug is added to cell culture media and is incorporated into the new strand during DNA replication in t ...
We describe a highly reproducible and multiplexed method, the GoldenGate assay for methylation, for high-throughput quantitative measurements of DNA methylation. It can analyze up to 1,536 targeted CpG sites in 96 samples simultaneously, using only 250 ng of genomic DNA. The method is akin ...
Restriction landmark genomic scanning (RLGS) is a method that provides a quantitative genetic and epigenetic (cytosine methylation) assessment of thousands of CpG islands in a single gel without prior knowledge of gene sequence. The method is based on two-dimensional separation of ra ...
Methylation-sensitive representational difference analysis (MS-RDA) is a genome subtraction method that isolates DNA fragments differentially methylated between two genomes. It can be performed in any organism, even in those for which no microarray products are available. An i ...
DNA methylation occurring on the 5 position of the pyrimidine ring of cytosines in the context of the dinucleotide sequence CpG forms one of the multiple layers of epigenetic mechanisms controlling and modulating gene expression through chromatin structure. It closely interacts with ...
Polyadenylation of nascent transcripts is an essential step for most mRNAs in eukaryotic cells. It is directly involved in the termination of transcription and is coupled with other steps of pre-mRNA processing. Recent studies have shown that transcript variants resulting from alter ...
The untranslated regions (UTRs) of many mRNAs contain sequence and structural motifs that are used to regulate the stability, localization, and translatability of the mRNA. It should be possible to discover previously unidentified RNA regulatory motifs by examining many related nuc ...
Riboswitches are intricate, metabolite-binding RNA structures found in the non-coding regions of mRNA. Allosteric changes in the riboswitch that are induced by metabolite binding are harnessed to control the genes of a variety of essential metabolic pathways in eubacteria and in some e ...
In eukaryotic organisms, gene regulatory networks require an additional level of coordination that links transcriptional and post-transcriptional processes. Messenger RNAs have traditionally been viewed as passive molecules in the pathway from transcription to transl ...
The regulation of mRNA turnover occurs in part through the action of mRNA-binding proteins that recognize specific nucleotide sequences and either activate or inhibit the decay of transcripts to which they are bound. In many cases, multiple mRNA-binding proteins, including those with o ...
Aptamers are an alternative to antibodies in their role as biorecognition elements in analytical devices. RNA aptamers, specific for different proteins, have been exploited as biorecognition elements to develop specific biosensors (aptasensors). These recognition elemen ...
RNA analysis by biosynthetic tagging (RABT) enables sensitive and specific queries of (a) how gene expression is regulated on a genome-wide scale and (b) transcriptional profiling of a single cell or tissue type in vivo. RABT can be achieved by exploiting unique properties of Toxoplasma gond ...
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