Accuracy and Reproducibility of Protein–DNA Microarray Technology
Identifieur interne : 002E57 ( Main/Exploration ); précédent : 002E56; suivant : 002E58Accuracy and Reproducibility of Protein–DNA Microarray Technology
Auteurs : Simon Field [Royaume-Uni] ; Irina Udalova [Royaume-Uni] ; Jiannis Ragoussis [Royaume-Uni]Source :
- Advances in Biochemical Engineering/Biotechnology [ 0724-6145 ]
Abstract
Abstract: Microarray-based methods for understanding protein–DNA interactions have been developed in the last 6 years due to the need to introduce high-throughput technologies in this field. Protein–DNA microarrays utilise chips upon which a large number of DNA sequences may be printed or synthesised. Any DNA-binding protein may then be interrogated by applying either purified sample or cellular/nuclear extracts, subject to availability of a suitable detection system. Protein is simply added to the microarray slide surface, which is then washed and subjected to at least one further incubation with a labelled molecule which binds specifically to the protein of interest. The signal obtained is proportional to the level of DNA-binding protein bound to each DNA feature, enabling relative affinities to be calculated. Key factors for reproducible and accurate quantification of protein binding are: microarray surface chemistry; length of oligonucleotides; position of the binding site sequence; quality of the protein and antibodies; and hybridisation conditions.
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DOI: 10.1007/10_2006_035
Affiliations:
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Protein–DNA microarrays utilise chips upon which a large number of DNA sequences may be printed or synthesised. Any DNA-binding protein may then be interrogated by applying either purified sample or cellular/nuclear extracts, subject to availability of a suitable detection system. Protein is simply added to the microarray slide surface, which is then washed and subjected to at least one further incubation with a labelled molecule which binds specifically to the protein of interest. The signal obtained is proportional to the level of DNA-binding protein bound to each DNA feature, enabling relative affinities to be calculated. Key factors for reproducible and accurate quantification of protein binding are: microarray surface chemistry; length of oligonucleotides; position of the binding site sequence; quality of the protein and antibodies; and hybridisation conditions.</div></front></TEI><affiliations><list><country><li>Royaume-Uni</li></country><region><li>Angleterre</li><li>Grand Londres</li><li>Oxfordshire</li></region><settlement><li>Londres</li><li>Oxford</li></settlement><orgName><li>Université d'Oxford</li></orgName></list><tree><country name="Royaume-Uni"><region name="Angleterre"><name sortKey="Field, Simon" sort="Field, Simon" uniqKey="Field S" first="Simon" last="Field">Simon Field</name></region><name sortKey="Ragoussis, Jiannis" sort="Ragoussis, Jiannis" uniqKey="Ragoussis J" first="Jiannis" last="Ragoussis">Jiannis Ragoussis</name><name sortKey="Ragoussis, Jiannis" sort="Ragoussis, Jiannis" uniqKey="Ragoussis J" first="Jiannis" last="Ragoussis">Jiannis Ragoussis</name><name sortKey="Udalova, Irina" sort="Udalova, Irina" uniqKey="Udalova I" first="Irina" last="Udalova">Irina Udalova</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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