Restriction Enzyme Treatment, Bisulfite Conversion, and Design of Bisulfite Sequencing Primers
1:54
Bisulfite Converted PCR and Gel Extraction
4:16
Bisulfite Sequencing Library Preparation
10:44
Results: Accurate Quantification of mtDNA Methylation
11:48
Conclusion
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The overall goal of this bisulfite sequencing protocol and bioinformatic pipeline is to achieve an accurate quantification of mitochondrial DNA methylation. This method can help answer key questions in the mitoepigenetic field such as is mitochond
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Here, we present a protocol to allow accurate quantification of mitochondrial DNA (mtDNA) methylation. In this protocol, we describe an enzymatic digestion of DNA with BamHI coupled with a bioinformatic analysis pipeline which can be used to avoid overestimation of mtDNA methylation levels caused by the secondary structure of mtDNA.