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It's because the bisulfite conversion procedure renders The 2 strands of DNA non-complementary, and in many situations primer design against one strand will deliver appropriate primers when the opposite strand will not likely.

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MSP-HTPrimer: a superior-throughput primer structure Instrument to enhance assay layout for DNA methylation Investigation in epigenetics

Bisulfite conversion of genomic DNA for methylation Assessment: protocol simplification with better Restoration applicable to confined samples and greater throughput

Bisulfite genomic sequencing will be the most generally made use of strategy to research the five-methylation of cytosines, the common covalent DNA modification in mammals. The procedure is based to the selective transformation of unmethylated cytosines to uridines. Then, the here investigated genomic regions are PCR amplified, subcloned and sequenced. During sequencing, the to begin with unmethylated cytosines are detected as thymines. The efficacy of bisulfite PCR is generally lower; mispriming and non-specific amplification frequently takes place as a result of T richness of your focus on sequences.

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