EZ DNA甲基化试剂盒
Format: Chemical denaturation of DNA followed
by sodium bisulfite modification, Spin Column.
? Recovery Volume: Purified,
bisulfite treated DNA is recovered in ≥ 10 μl provided elution buffer.
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Input DNA: 500 pg - 2 μg DNA per treatment with 200 - 500 ng being optimal.
? Sample Sources: Plasmid, genomic, and endonuclease digested DNAs.
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Equipment: Microcentrifuge
Descriptlon
DNA Methylation is a
naturally occurring event that happens in both prokaryotic and eukaryotic
organisms. In prokaryotes DNA methylation provides a way to protect host DNA
from digestion by their own restriction enzymes that are designed to eliminate
foreign DNA. In higher eukaryotes DNA methylation acts as another level of gene
regulation. It has been clearly demonstrated that aberrant methylation is a
widespread phenomenon in cancer and may be among the earliest changes during
oncogenesis. DNA methylation has also been shown to play a central role in gene
imprinting, embryonic development, X-Chromosome gene silencing and cell cycle
regulation. In many plants and animals, including mammals, DNA methylation
consists of the addition of a methyl group to the fifth carbon position of the
cytosine pyrimidine ring via a methyltransferase enzyme. The majority of the DNA
methylation in mammals is found in 5’-CpG-3’ dinucleotides, but other
methylation patterns do exist. In fact, about 80 percent of all 5’-CpG-3’
dinucleotides in mammalian genomes are methylated, and the majority of the 20
percent that remain unmethylated are found within promoters or in the first
exons of genes. It is obvious that the ability to quantify and detect DNA
methylation efficiently and accurately is essential for the study of cancer,
gene expression, genetic diseases, and many other important aspects of biology.
To date, a number of methods have been developed to quantify DNA methylation
such as high performance capillary electrophoresis and methylation-sensitive
arbitrarily primed PCR, but currently the most commonly used technique is the
bisulfite method. This technique consists of treating DNA with bisulfite, which
causes unmethylated cytosines to be converted into uracil while methylated
cytosines remain unchanged. In this protocol the bisulfite modified DNA is
amplified by PCR and the resulting PCR products are either analyzed by DNA
sequencing or restriction endonuclease digestion. The methylation status of the
DNA segment is then determined by comparing the bisulfite treated DNA to the
untreated DNA.
The EZ DNA Methylation? Kit uses a simplified procedure
and streamlines the bisulfite method for DNA methylation analysis. The kit is
based on the three step reaction that takes place between cytosine and sodium
bisulfite where cytosine is converted into uracil. The EZ DNA Methylation Kit’s
innovative in-column desulphonation reaction eliminates several precipitation
steps and provides researchers with consistent reaction conditions. The kit is
designed to reduce template degradation, minimize DNA loss during treatment and
clean-up, and result in the complete conversion of unmethylated cytosines. The
recovered, treated DNA template is ideal for Methylation Specific PCR (MSP)
followed by methylation analysis using restriction endonucleases, sequencing,
microarrays, etc.
Product Catalog No. Size
EZ DNA
Methylation? Kit D5001 50 Rxns.