DNA Footprinting Assay Kit
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.
Kit Components:
10x FP Buffer: 1 ml, store at -20ºC
0.1 mg/ml poly(dI-dC): 0.5 ml, store at -20ºC (100-500 assays)
DNase: 0.1 ml, store at -20ºC
FP Loading Solution: 1 ml, store at -20ºC (contains 90% formamide)
DNase Stop Solution: 10 ml, store at 4ºC
DNA Phenol: 10 ml, store at 4ºC (contains 50% phenol and 50% chloroform)
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.
Kit Components:
10x FP Buffer: 1 ml, store at -20ºC
0.1 mg/ml poly(dI-dC): 0.5 ml, store at -20ºC (100-500 assays)
DNase: 0.1 ml, store at -20ºC
FP Loading Solution: 1 ml, store at -20ºC (contains 90% formamide)
DNase Stop Solution: 10 ml, store at 4ºC
DNA Phenol: 10 ml, store at 4ºC (contains 50% phenol and 50% chloroform)
Biomedical Research Service
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