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Epigenomics: ChIP-Seq, DNase-Seq, FAIRE, ATAC-Seq, Nucleosome positioning

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Modified: 26 March 2020, 12:24 PM   User: Emilia Petrachi  → Emilia Petrachi

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DNase-Seq

(Emilia Petrachi)

DNase-Seq is one of the several approaches in molecular biology useful to identify DNA response elements, or regulatory regions in general, through genome-wide sequencing of regions sensitive to cleavage by DNase I.
A brief outline of the technique is the following:

  1. DNA-protein complexes are treated with DNase I;
  2. DNA extraction and sequencing are perfomed;
  3. Sequences bound by regulatory proteins are protected  from DNase I digestion;
  4. Deep sequencing is performed to provide accurate representation of location of regulatory proteins in the genome.

Pros

  • Can detect open chromatin
  • No prior knowledge of the sequence or binding protein is required
  • Compared to formaldehyde-assisted isolation of regulatory elements and sequencing (FAIRE-seq), has greater sensitivity at promoters

Cons

  • DNase l is sequence-specific and hypersensitive sites might not account for the entire genome
  • DNA loss through the multiple purification steps limits sensitivity
  • Integration of DNase I with ChIP data is necessary to identify and differentiate similar protein-binding sites

More information can be found at the website:
https://emea.illumina.com/science/sequencing-method-explorer/kits-and-arrays/dnase-seq-dnasel-seq.html
And this is a video made by a Biology Professor at Davidson College, it explains the protocol in really easy terms:

Brief outline of the DNase-Seq protocol

Another outline of the protocol

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