BisulfiteSequencingofDNA RussellP.Darst,CarolinaE.Pardo,LingbaoAi,KevinD.Brown,andMichaelP.Kladde * DepartmentofBiochemistryandMolecularBiologyandUFShandsCancerCenterProgramin CancerGenetics,EpigeneticsandTumorVirology,UniversityofFloridaCollegeofMedicine
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Bisulfite modification of DNA is the most commonly used, "gold standard" method for DNA methylation studies. Since bisulfite treatment introduces specific changes in the DNA sequence depending on the methylation status of individual cytosine residues
current protocols in molecular biology volume 3 || bisulfite sequencing of dnadoi:10.1002/0471142727Molecular biologyThis compendium of techniques coverage of state-of-the-art developments in molecular biology, with over 600 pages of information contributed by a wide range of author...
The biological roles of DNA methylation have been elucidated by profiling methods based on whole-genome or reduced-representation bisulfite sequencing, but these approaches do not efficiently survey the vast numbers of non-coding regulatory elements in mammalian genomes. Here we present an extended-repre...
Bisulfite-seq is the golden standard for DNA methylation analysis. Process includes: treating of DNA with bisulfite to convert cytosine residues to uracil, while leaving 5-methylcytosine residues unaffected; running PCR to convert all the uracil to thymines; in the end, sequencing the PCR product ...
Figure 1. Bisulfite conversion and sequencing of samples (Li, 2011) Methylation on a DNA strand could be detected when unmethylated cytosines in single-stranded DNA are converted into uracil residues which are recognized as thymine in subsequent PCR amplification. On the other hand, methylated c...
Here we demonstrate a droplet-based microfluidic technology, Drop-BS, to construct single-cell bisulfite sequencing libraries for DNA methylome profiling. Drop-BS takes advantage of the ultrahigh throughput offered by droplet microfluidics to prepare bisulfite sequencing libraries of up to 10,000 single...
DNA MethylationEpigeneticsCancerThe bisulfite genomic sequencing (BGS) protocol has gained worldwide popularity as the method of choice for analyzing DNA methylation. It is this popular because it is a powerful protocol and it may be coupled with many other applications. However, users often run ...
An accurate and reproducible next-generation sequencing platform is essential to identify malignancy-related abnormal DNA methylation changes and translate them into clinical applications including cancer detection, prognosis, and surveillance. However,