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Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma
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JoVE Journal Genetics
Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma

Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma

DOI:
10.3791/54399-v

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08:53 min

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June 10, 2017

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, , , , ,
1Innovation Laboratory, Center for Molecular Oncology,Memorial Sloan Kettering Cancer Center, 2Flow-Sorting Core Facility,Weill Cornell Medical College, 3Department of Laboratory Medicine,University of Washington, 4Department of Physiology and Biophysics, Institute for Computational Biomedicine,Weill Cornell Medical College, 5Department of Pathology and Laboratory Medicine,Weill Cornell Medical College, 6Department of Laboratory Medicine,Memorial Sloan Kettering Cancer Center

Chapters

  • 00:05Title
  • 01:19Cell Suspension Preparation for Sorting
  • 03:00Compensation Control Setup
  • 04:53Hodgkin Reed-Sternberg-, B-, and T-cell Isolation
  • 06:07Results: Representative DNA Library and Copy Number Gains and Losses Analyses
  • 08:02Conclusion

Summary

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Here, we describe a combined flow cytometric cell sorting and low-input, next-generation library construction protocol designed to produce high-quality, whole-exome data from the Hodgkin Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (CHL).

Tags

Keywords: Flow-sortingExome SequencingReed-Sternberg CellsClassical Hodgkin LymphomaCHLHRS CellsNext-generation SequencingTumor-derived DNADiagnosisClassificationPrognosisTherapyFlow CytometryAntibody CocktailThawing MediumSorting Medium

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