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Reusable Single Cell for Iterative Epigenomic Analyses
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JoVE 杂志 遗传学
Reusable Single Cell for Iterative Epigenomic Analyses

Reusable Single Cell for Iterative Epigenomic Analyses

DOI:
10.3791/63456-v

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10:28 min

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February 11, 2022

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, , ,
1Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute,National Institutes of Health, 2Biostatistics and Data Management Section, Center for Cancer Research, National Cancer Institute,National Institutes of Health, 3CCR Collaborative Bioinformatics Resource (CCBR), Center for Cancer Research, National Cancer Institute,National Institutes of Health, 4Advanced Biomedical Computational Science,Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute

Chapters

  • 00:05Introduction
  • 01:32Preparation of Reusable Single Cells: Manual Version
  • 03:09Random Primer Annealing and Extension
  • 04:09Antibody Binding and Proximity Ligation of Antibody Probe and Proximally Extended Random Primer
  • 05:25Full Extension of the 1st Random Primer and Multiple Displacement Amplification
  • 07:32Results: Single Cell Multiepigenomic Analysis
  • 09:26Conclusion

Summary

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The present protocol describes a single-cell method for iterative epigenomic analyses using a reusable single cell. The reusable single cell allows analyses of multiple epigenetic marks in the same single cell and statistical validation of the results.

Tags

Reusable Single CellEpigenomic AnalysisSingle-cell EpigenomeEpigenetic TherapyEpigenome RegulationGene ExpressionIntercalator DyeEDTA PBSPCR TubeSwing RotorLiquid Handling RobotTEMEDMineral OilTP Magnesium Negative BufferAnnealing Buffer

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