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Chromatin Interaction Analysis gepaart mit End-Tag-Sequenzierung (Chia-PET) zur Abbildung von Chromatin-Wechselwirkungen zu verstehen und Transkriptionsregulation
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JoVE 신문 생물학
Chromatin Interaction Analysis with Paired-End Tag Sequencing (ChIA-PET) for Mapping Chromatin Interactions and Understanding Transcription Regulation
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Chromatin Interaction Analysis gepaart mit End-Tag-Sequenzierung (Chia-PET) zur Abbildung von Chromatin-Wechselwirkungen zu verstehen und Transkriptionsregulation

DOI:
10.3791/3770-v

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21:55 min

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April 30, 2012

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, , , , , , ,
1Genome Institute of Singapore,Agency for Science, Technology and Research, Singapore, 2A*STAR-Duke-NUS Neuroscience Research Partnership, Singapore, 3Department of Biochemistry,National University of Singapore, Singapore

Chapters

  • 00:02Title
  • 01:00Overall View of Chromatin Interaction Analysis using Paired-end Tag (ChIA-PET)
  • 02:15ChIP – Chromatin Immunoprecipitation
  • 04:24Cell Lysis
  • 05:07Nuclear Lysis
  • 07:11Fragmentation of Chromatin
  • 08:26Washing, Preclearing, and Coating of Antibody to Beads
  • 10:08Chromatin Immunopreciptation
  • 11:20Washing and Elution
  • 11:48ChIA-PET – Chromatin Interaction Analysis using Paired-end Tag Sequencing
  • 12:29End-blunting of ChIP DNA Fragments
  • 13:33Ligation of Biotinylated Half-linkers to ChIP DNA
  • 16:02Elution and Proximity Ligation of ChIP DNA Fragments
  • 18:08Immobilization of ChIA-PET DNA to Streptavidin Beads
  • 20:47Conclusion

Summary

자동 번역

자동 번역

Chromatin Interaction Analysis von Paired-End-Tag-Sequenzierung (Chia-PET) ist eine Methode zur<em> De-novo-</em> Nachweis von Chromatin-Wechselwirkungen, zum besseren Verständnis der transkriptionellen Kontrolle.

Tags

Chromatin Interaction AnalysisPaired-End Tag SequencingChIA-PETMapping Chromatin InteractionsTranscription RegulationThree-dimensional StructuresHigher-order ConformationsNuclear SpacesGene PromotersRegulatory ElementsTranscription FactorsRegulatory SequencesNetwork Of Transcription RegulationCoordinationChromatin ImmunoprecipitationChIPChromatin FragmentsProximity LigationCovalent LinksDNA FragmentsChromatin ComplexesMmeI Restriction Enzyme SitesHalf-linkersPET Constructs

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