A simple method for DNaseI footprinting analysis.
DNaseI footprinting technique has been a very useful method in gaining direct and immediate information about the location of a protein binding site in the DNA sequence. In this report, we present a method that overcomes many of the inconveniences of previous methods. This method was based on the combination of PCR technique and the dideoxy DNA sequencing reaction. It avoids the need of the secondary restriction sites that any candidate DNA sequence for DNaseI footprinting analysis can be prepared efficiently as long as its flanking sequences are known. Replacing the difficult Maxam-Gilbert DNA sequencing with the dideoxy-mediated DNA sequencing method makes the footprinting experiments even safer and easier.
Lin
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Department of Plant Pathology, University of Minnesota, Minneapolis 55414, USA.
Shiuan
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D001483:Base Sequence; D001665:Binding Sites; D016172:DNA Fingerprinting / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000737:chemistry*; D016133:Polymerase Chain Reaction
D004268:DNA-Binding Proteins; D003850:Deoxyribonuclease I