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DNA Footprinting Assay Kit

Components:

            10x FP Buffer: 1 ml, store at -20ºC

            0.1 mg/ml poly(dI-dC): 0.5 ml, store at -20ºC

            DNase: 0.1 ml, store at -20ºC

            FP Loading Solution: 1 ml, store at -20ºC

            DNase Stop Solution: 10 ml, store at 4ºC

            DNA Phenol: 10 ml, store at 4ºC

Product Description: DNAase I in the presence of Mg²⁺ hydrolyzes each strand of DNA independently in a statistically random fashion. This property of the nuclease proves to be quite useful in the analysis of DNA-protein complexes (in a technique commonly referred to as DNA footprinting assay; Nucl. Acids Res. 5:3157,1978) since the protein-bound region is often protected from the attack of properly diluted DNAase I. Information regarding structural features of DNA such as the distribution of DNAase I hyper-sensitive sites within the DNA segment of interest is also readily obtainable. The footprinting assay, which is best complemented by gel shift assay, can be performed with purified protein factors or with a crude cell extract in the presence of a non-specific polyanion competitor, usually double-stranded poly(dI-dC). The amount of poly(dI-dC) used per reaction can be much reduced if purified protein fractions are used for the analysis of DNA-protein complexes. The product is stable for at least 1 year if stored and handled properly, and is for research use only.

Features: Components are optimized for footprinting assays using either crude cell extracts or purified protein fractions.