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Adipocyte-Specific ATAC-Seq with Adipose Tissues Using Fluorescence-Activated Nucleus Sorting
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JoVE 杂志 遗传学
Adipocyte-Specific ATAC-Seq with Adipose Tissues Using Fluorescence-Activated Nucleus Sorting

Adipocyte-Specific ATAC-Seq with Adipose Tissues Using Fluorescence-Activated Nucleus Sorting

DOI:
10.3791/65033-v

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11:11 min

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March 17, 2023

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, , , ,
1Department of Biochemistry and Molecular Biology,Indiana University School of Medicine

Chapters

  • 00:05Introduction
  • 00:51Nucleus Isolation
  • 02:15Nucleus Sorting and Tn5 Tagmentation
  • 04:08DNA Purification and PCR Amplification
  • 05:32Real-Time Quantitative PCR (qPCR) Test, Additional PCR Amplification, and Second DNA Purification
  • 06:14DNA Fragment Size Selection
  • 08:01Results: Assay for Transposase-Accessible Chromatin with High-Throughput Sequencing
  • 10:19Conclusion

Summary

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We present a protocol for assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) specifically on adipocytes using nucleus sorting with adipose tissues isolated from transgenic reporter mice with nuclear fluorescence labeling.

Tags

Adipocyte-specific ATAC-seqFluorescence-activated Nucleus SortingGene Expression Regulatory MechanismsNuclear IsolationFACSTn5 TransposaseATAC-sequencing

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