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• W2034793893 endingPage "3209" @default.
• W2034793893 startingPage "3203" @default.
• W2034793893 abstract "We developed a rapid method designated Target Detection Assay (TDA) to determine DNA binding sites for putative DNA binding proteins. A purified, functionally active DNA binding protein and a pool of random double-stranded oligonucleotides harbouring PCR primer sites at each end are included the TDA cycle which consists of four separate steps: a DNA protein incubation step, a protein DNA complex separation step, a DNA elution step and a polymerase chain reaction (PCR) DNA amplification step. The stringency of selection can be increased in consecutive TDA cycles. Since tiny amounts of retained DNA can be rescued by PCR, buffer systems, salt concentrations and competitor DNA contents can be varied in order to determine high affinity binding sites for the protein of choice. To test the efficiency of the TDA procedure potential DNA binding sites were selected by the DNA binding protein SP1 from a pool of oligonucleotides with random nucleotides at 12 positions. Target sites selected by recombinant SP1 closely matched the SP1 consensus site. If DNA recognition sites have to be determined for known, mutated or putative DNA binding proteins, the Target Detection Assay (TDA) is a versatile and rapid technique for consideration." @default.
• W2034793893 created "2016-06-24" @default.
• W2034793893 creator A5040100465 @default.
• W2034793893 creator A5061531129 @default.
• W2034793893 date "1990-01-01" @default.
• W2034793893 modified "2023-10-13" @default.
• W2034793893 title "Target Detection Assay (TDA): a versatile procedure to determine DNA blinding sites as demonstrated on SP1 protein" @default.
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• W2034793893 doi "https://doi.org/10.1093/nar/18.11.3203" @default.
• W2034793893 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/330924" @default.
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