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Bisulfite Conversion
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Bisulfite Conversion is a process in which double-stranded genomic DNA is treated with sodium bisulfite, leading to deamination of unmethylated cytosines into uracils, while methylated cytosines remain unchanged. The DNA is then amplified by PCR with primers that differentiate between methylated and unmethylated sequences followed by sequencing analysis. A comparison of the sequences of converted and untreated DNA will reveal the methylation profile of the sample. The results provide single nucleotide resolution information about the methylation status of a particular region of DNA. Alternatively, non-sequencing methods can be used to determine the methylation status at a genome-wide level.
Active Motif's MethylDetector™ Bisulfite Modification Kit simplifies analysis of DNA methylation. It comes complete with optimized reagents for performing DNA conversion with bisulfite, plus time-saving DNA purification columns and positive control PCR primers to validate your results.
In the MethylDetector method, DNA of interest is rapidly heat denatured in a thermocycler in the presence of the bisulfite conversion reagent. The temperature is then lowered and the conversion reaction is performed. Unlike other methods, MethylDetector does not require an initial acid denaturation step as the conversion reagent includes a DNA denaturant, which saves you time and effort. After DNA conversion, the sample is added to the included DNA purification columns, and a simple, on-column desulfonation is performed. Ready-to-use DNA is then eluted from the columns. For your convenience, the included positive control PCR primers can be used to assess the success of the bisulfite conversion before you spend time and money on DNA sequencing. This is because the included primers only anneal to converted human DNA
Additionally, for those researchers interested in Bisulfite Sequencing Services, Active Motif now offers bisulfite sequencing as a custom service. The sequencing service includes primer design, isolation of DNA from cells, bisulfite conversion of DNA, PCR amplification and gel analysis of PCR products, gel purification of PCR products, cloning of PCR products, amplification of cloned inserts, sequencing and data analysis. Click to learn more about our Bisulfite Sequencing Services.
