Methylation Analysis by Restriction Endonuclease Digestion and Real-Time PCR
نویسندگان
چکیده
منابع مشابه
Methylation analysis by restriction endonuclease digestion and real-time PCR.
DNA methylation is a highly characterized epigenetic modification of the human genome that is critical for normal cell processes, including tissue regulation, genetic expression development, genomic imprinting, X-chromosome inactivation, and DNA repair. DNA methylation in humans occurs almost exclusively at the C5 position of CpG dinucleotides in genomic promoter regions. Alterations in DNA met...
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The polymerase chain reaction (PCR) has become the most widely used technique for the manipulation of DNA with applications for cDNA cloning, gene cloning, polymorphism detection, mutagenesis and allele-specific diagnosis (8). All of those applications have been extended significantly by the recent development of “long” PCR, a technique that makes it possible to amplify DNA fragments up to 40 k...
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One of the common methods for cloning polymerase chain reaction (PCR) products is overhanging-end cloning (also known as sticky-end or directional cloning). Frequently, it is not possible to use restriction enzyme sites already present in the amplified product, and primers that encode recognition sites of restriction endonucleases in addition to the specific sequence have to be designed. After ...
متن کاملRecognition of native DNA methylation by the PvuII restriction endonuclease.
Recognizing the methylation status of specific DNA sequences is central to the function of many systems in eukaryotes and prokaryotes. Restriction-modification systems have to distinguish between 'self' and 'non-self' DNA and depend on the inability of restriction endonucleases to cleave their DNA substrates when the DNA is appropriately methylated. These endonucleases thus provide a model syst...
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BACKGROUND Epigenetic studies, such as the measurement of DNA methylation, are important in the investigation of syndromes influenced by imprinted genes. Quick and accurate quantification of methylation at such genes can be of appreciable diagnostic aid. METHODS We first digested genomic DNA with methylation-sensitive restriction enzymes and used DNA without digestion as a control and nonmeth...
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ژورنال
عنوان ژورنال: Clinical Chemistry
سال: 2010
ISSN: 0009-9147,1530-8561
DOI: 10.1373/clinchem.2010.146654