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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

DOI:
10.3791/52246-v

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13:47 min

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February 24, 2015

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, , , , , , , ,
1Department of Medicine,Weill Cornell Medical College, 2Institute for Computational Biomedicine,Weill Cornell Medical College, 3Department of Physiology and Biophysics,Weill Cornell Medical College, 4Department of Pathology,University of Michigan

Chapters

  • 00:05Title
  • 02:22Adapter Ligation and DNA Size-selection via Gel Purification
  • 07:32Bisulfite Conversion, Enrichment PCR, and DNA Purification
  • 09:40Library Preparation for Sequencing
  • 10:42Results: ERRBS Yields Base Pair Resolution CpG Methylation Data
  • 12:15Conclusion

Summary

Automatic Translation

Automatic Translation

Enhanced Reduced Representation Bisulfite Sequencing is a method for the preparation of sequencing libraries for DNA methylation analysis based on restriction enzyme digestion combined with cytosine bisulfite conversion. This protocol requires 50 ng of starting material and yields base pair resolution data at GC-rich genomic regions.

Tags

Keywords: DNA MethylationBisulfite SequencingReduced RepresentationERRBSBase Pair ResolutionQuantitative MethylationGenomic LociRestriction EnzymeNext-generation SequencingBioinformatics Analysis

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