11/16/2023 0 Comments Touchdown pcr bisulfite sequencingIt has been shown to have an important role in the transcriptional regulation of multiple physiological processes. Accordingly, this novel method is anticipated to be an efficient and economical alternative tool for rapid quantification of methylation patterns in screening large numbers of clinical samples across multiple genes.ĭNA methylation at the carbon-5 position of the cytosine base in CpG dinucleotides is a common chemical modification of DNA in eukaryotes. In comparison with pyrosequencing and bisulfite-cloning sequencing, our method is confirmed to be a simple, high-throughput and cost-effective technology for determining the methylation status of specific genes. Furthermore, we take several effective measures to break through the ‘bottleneck’ problems that render the routine bisulfite sequencing method unsuitable for quantitative methylation. According to the principles of bisulfite-PCR, converting unmethylated cytosines to thymine while leaving methylated cytosines unchanged, we regard the CpG site as a SNP and estimate the methylation status of cytosines in the given CG dinucleotides by measuring the ratio of the cytosine peak height to the sum of cytosine and thymine peak heights in automated DNA sequencing traces. In this report, we describe a novel method for the rapid quantification of CpG methylation on the basis of direct bisulfite-PCR sequencing method. ![]() However, rapid and simple methods for determining the DNA methylation status of CpG sites in genes still remain elusive. Various technologies are currently available to quantify DNA methylation.
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