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Determine DNase hyper sensitive site in the promoter region

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DNase hyper sensitive sites are important for changing chromatin structure during transcriptional activation of genes.

Use BOTH diagram & written explanation:

Determine whether DNase (HSS) formation at specific sites in promoter regions is an important component of transcriptional activation of a gene you are studying during your research.

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The solution gives the detailed information and diagrams as to how to determine the DNase hyper sensitive site in the promoter region for the transcriptional activation of genes.

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For DNAase sensitive site analysis, first prepare enough each cells (around 107 cells) from several cell lines. From these cells, prepare nuclear suspension by lysing these cells and removing cell pellets. For hypersensitive sites, nuclear suspension is mixed with various concentration of (0-560 U.ml-1) DNAase I for 10 min on ice. The reaction is stopped by EDTA and treated with proteinase K. After ...

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