Student Wiki on methodology
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Epigenomics: ChIP-Seq, DNase-Seq, FAIRE, ATAC-Seq, Nucleosome positioning
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Modified: 26 March 2020, 12:21 PM User: Emilia Petrachi →
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Epigenomics:
ChIP-Seq
DNase-Seq
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:
- DNA-protein complexes are treated with DNase I;
- DNA extraction and sequencing are perfomed;
- Sequences bound by regulatory proteins are protected from DNase I digestion;
- 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:
(Emilia Petrachi)