| Literature DB >> 19378165 |
Abstract
A technique is described for the identification of nucleic acid sequences bound with high affinity by proteins or by other molecules suitable for a partitioning assay. Here, a histidine-tagged protein is allowed to interact with a pool of nucleic acids and the protein-nucleic acid complexes formed are retained on a Ni-NTA matrix. Nucleic acids with a low level of recognition by the protein are washed away. The pool of recovered nucleic acids is amplified by the polymerase chain reaction and is submitted to further rounds of selection. Each round of selection increases the proportion of sequences that are avidly bound by the protein of interest. The cloning and sequencing of these sequences finally completes their identification.Entities:
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Year: 2009 PMID: 19378165 DOI: 10.1007/978-1-60327-015-1_11
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745