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Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients
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Genetics
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JoVE Journal Genetics
Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients

Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients

DOI:
10.3791/55773-v

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13:21 min

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June 16, 2017

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1Department of Medical Genetics, Institute of Biomedicine,Gothenburg University, 2Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine,Gothenburg University

Chapters

  • 00:05Title
  • 00:49Genomic DNA Extraction and Sonication
  • 02:06Bead Preparation Prior to Binding with Methyl-CpG-binding Domain (MBD)-biotin Protein
  • 03:35Binding the MBD-biotin Protein to the Washed Beads
  • 04:59Binding MBD-biotin Beads with Fragmented Genomic DNA
  • 06:22Removing the Unbound DNA and Eluting the Methylated DNA from the Beads
  • 08:10Ethanol Precipitation and the Enrichment of Methylated DNA
  • 10:09Results: Identification of Differentially Methylated CpG Rich Regions in Chronic Lymphocytic Leukemia (CLL) Samples
  • 11:26Conclusion

Summary

Automatic Translation

Automatic Translation

This work describes an optimized methyl-CpG-binding domain (MBD) sequencing protocol and a computational pipeline to identify differentially methylated CpG-rich regions in chronic lymphocytic leukemia (CLL) patients.

Tags

Keywords: DNA MethylationMethyl-CpG-binding DomainCapture-based MethodChronic Lymphocytic LeukemiaNext-generation SequencingGlobal Methylation ProfilingDifferentially Methylated CpG-rich RegionsCLL-specific Signature GenesDisease PathogenesisPrognosisGenomic DNA ExtractionSonicationMagnetic Streptavidin BeadsMBD-biotin Protein

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