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Downloaded from genome.ship.org on May 20, 2015, Published by Cold Spring Harbor Laboratory Press Methods Genome-wide, high-resolution DNA methylation profiling using bisulfite-mediated cytosine conversion

How to fill out genome-wide high-resolution dna methylation

How to fill out genome-wide high-resolution DNA methylation:

1. Obtain the DNA samples: Collect biological samples, such as blood or tissue, from the individuals or organisms of interest. Ensure proper handling and storage to preserve DNA integrity.
2. DNA extraction: Isolate the DNA from the collected samples using appropriate extraction methods. This step separates the DNA from other cellular components.
3. Bisulfite conversion: Perform bisulfite conversion on the extracted DNA. This chemical treatment converts unmethylated cytosines to uracil, while leaving methylated cytosines unchanged. This allows the differentiation between methylated and unmethylated regions of DNA.
4. Library preparation: Generate libraries from the bisulfite-converted DNA samples. This involves fragmenting the DNA, adding adapters, and amplifying specific regions of interest.
5. Sequencing: Perform high-resolution sequencing on the prepared libraries. Next-generation sequencing technologies, such as whole-genome bisulfite sequencing or reduced representation bisulfite sequencing, can provide genome-wide coverage of DNA methylation patterns.
6. Data analysis: Analyze the sequencing data to determine DNA methylation patterns across the genome. This involves aligning sequencing reads to a reference genome, identifying differentially methylated regions, and quantifying methylation levels at specific loci.
7. Interpretation: Interpret the obtained DNA methylation data in the context of the research question or clinical application. Compare the methylation patterns between different samples or conditions to understand their potential functional implications.
8. Reporting and dissemination: Document the findings and communicate the results through scientific publications, reports, or presentations to contribute to the field's knowledge.

Who needs genome-wide high-resolution DNA methylation?

1. Researchers studying epigenetics: Genome-wide high-resolution DNA methylation analysis is valuable to researchers interested in understanding the role of DNA methylation in gene regulation, development, disease progression, or environmental influences.
2. Biomedical scientists: Genome-wide DNA methylation profiling can aid in identifying epigenetic changes associated with various diseases, such as cancer, neurological disorders, or autoimmune conditions. This information can potentially contribute to the development of diagnostic biomarkers or therapeutic targets.
3. Personalized medicine practitioners: By analyzing genome-wide DNA methylation patterns, healthcare professionals may gain insights into an individual's susceptibility to certain diseases or predict treatment response. This could aid in tailoring personalized treatment plans or interventions.

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