Why is target dna treated with sodium bisulphite?
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Bisulfite-Conversion-Based Assays
Sodium bisulfite DNA treatment allows for discrimination between methylated and unmethylated cytosines. Bisulfite conversion was one of the first techniques developed for methylation analysis and is still considered the gold standard [21] because it allows for quantitative comparisons of methylation levels at single-base resoluttttion
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Sodium bisulfite DNA treatment allows for discrimination between methylated and unmethylated cytosines. Bisulfite conversion was one of the first techniques developed for methylation analysis and is still considered the gold standard [21] because it allows for quantitative comparisons of methylation levels at single-base resoluttttion
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sodium bisulfite is used in the analysis of themethylation status of cytosines in DNA.
In this technique, sodium bisulfite deaminatescytosine into uracil, but does not affect 5-methylcytosine, a methylated form of cytosine with a methyl group attached to carbon 5.
When the bisulfite-treated DNA is amplified via polymerase chain reaction, the uracil is amplified as thymine and the methylated cytosines are amplified as cytosine. DNA sequencing techniques are then used to read the sequence of the bisulfite-treated DNA. Those cytosines that are read as cytosines after sequencing represent methylated cytosines, while those that are read as thymines represent unmethylated cytosines in the genomic DNA.
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In this technique, sodium bisulfite deaminatescytosine into uracil, but does not affect 5-methylcytosine, a methylated form of cytosine with a methyl group attached to carbon 5.
When the bisulfite-treated DNA is amplified via polymerase chain reaction, the uracil is amplified as thymine and the methylated cytosines are amplified as cytosine. DNA sequencing techniques are then used to read the sequence of the bisulfite-treated DNA. Those cytosines that are read as cytosines after sequencing represent methylated cytosines, while those that are read as thymines represent unmethylated cytosines in the genomic DNA.
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