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Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique
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Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique

Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique

DOI:
10.3791/53647-v

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09:14 min

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January 14, 2016

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1Department of Radiation Oncology, Department of Oncological Sciences, Huntsman Cancer Institute,University of Utah School of Medicine

Chapters

  • 00:05Title
  • 00:44Chromatin Preparation with BrdU Labeling
  • 03:06Chromatin Immunoprecipitation (ChIP)
  • 05:28Slot Blot Analysis of Chip DNA
  • 07:22Results: Loss of Histone Deacetylase 1 and 2 Increases H4K16ac on Nascent DNA and SMARCA5 Associates with Nascent DNA
  • 08:21Conclusion

Summary

자동 번역

자동 번역

In this protocol, we describe a novel BrdU-ChIP-Slot-Western technique to examine proteins and histone modifications associated with newly synthesized or nascent DNA.

Tags

Proteins BoundNascent DNABrdU-ChIP-Slot-Western TechniqueBrdU LabelingChromatin PrecipitationReplicationRepairProteinsDNA ReplicationReplication StressAdvantageQuick MethodQuantitative MannerNIH 3T3 CellsMediaDMSOInhibitorsBrdU ConcentrationIncubateCrosslinkFormaldehydeConcentrationRoom TemperatureAgitationQuench ReactionGlycinePBSCentrifugationPelletFrozen StorageLysate Buffer

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