EpiMark® Bisulfite Conversion Kit
Bisulfite conversion is the most commonly used technique to date and is the “gold standard” for methylation analysis.
- Incubation of the target DNA with sodium bisulfite results in conversion of all unmodified cytosines to uracils leaving the modified bases (5-mC or 5-hmC) intact
- Complete conversion of Cytosine is achieved by alternating cycles of thermal denaturation with incubation reactions
- This kit provides materials for 48 reactions
Genomic DNA from many organisms has modified nucleotides. In the mammalian genome, the modified base is predominately 5-methylcytosine (5-mC), which is involved in gene expression regulation, including selective inactivation of one X chromosome in females of mammalian species (1,2). Symmetrical CpG DNA methylation is heritable, but also reversible. This change of methylation status results in an enormous number of combinations of epigenetic states that can regulate gene expression. In mammalian cells, DNA methylation mainly occurs in CpG dinucleotides and is carried out by two methyltransferase enzymatic activities, namely, maintenance methylation and de novo methylation. The maintenance methyltransferase, DNMT1, is involved in the DNA methylation after every cellular DNA replication cycle. DNMT3a and DNMT3b are the de novo methyltransferases that are active in early development.
For optimal results, 50 ng–2 μg DNA is recommended.
Each EpiMark Bisulfite Conversion Kit contains sufficient reagents to perform the bisulfite conversion reaction of 48 samples. Store at room temperature (15–20°C). However, for long-term storage (longer than 4 weeks) Desulphonation Reaction Buffer concentrate should be stored at 2–8°C. Prepared Bisulfite Mix can be stored at -20°C for up to six months.
The following reagents are supplied with this product:
Store at (°C) Concentration Bisulfite Mix 25 Solubilization Buffer Desulfonation Buffer (concentrate) 25 10 X Binding Buffer 25 1 X Wash Buffer (concentrate) 25 5 X Elution Buffer 25 1 X EpiMark spin columns with 2 ml collection tubes 50
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