共查询到20条相似文献,搜索用时 15 毫秒
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M Garber N Yosef A Goren R Raychowdhury A Thielke M Guttman J Robinson B Minie N Chevrier Z Itzhaki R Blecher-Gonen C Bornstein D Amann-Zalcenstein A Weiner D Friedrich J Meldrim O Ram C Cheng A Gnirke S Fisher N Friedman B Wong BE Bernstein C Nusbaum N Hacohen A Regev I Amit 《Molecular cell》2012,47(5):810-822
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A new analytical scale DNA affinity binding assay for analyses of specific protein-DNA interactions.
We describe a rapid analytical assay for identification of proteins binding to specific DNA sequences. The DAPSTER assay (DNA affinity preincubation specificity test of recognition assay) is a DNA affinity chromatography-based microassay that can discriminate between specific and nonspecific protein-DNA interactions. The assay is sensitive and can detect protein-DNA interactions and larger multicomponent complexes that can be missed by other analytical methods. Here we describe in detail the optimization and utilization of the DAPSTER assay to isolate AP-1 complexes and associated proteins in multimeric complexes bound to the AP-1 DNA element. 相似文献
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Discrimination of base mismatches from normal Watson-Crick base pairs in duplex DNA constitutes a key approach to the detection of single nucleotide polymorphisms (SNPs). We have developed a sensor for a surface plasmon resonance (SPR) assay system to detect G-G, A-A, and C-C mismatch duplexes by employing a surface upon which mismatch-binding ligands (MBLs) are immobilized. We synthesized a new MBL consisting of 2,7-diamino-1,8-naphthyridine (damND) and immobilized it onto a CM5 sensor chip to carry out the SPR assay of DNA duplexes containing a single-base mismatch. The SPR sensor with damND revealed strong responses to all C-C mismatches, and sequence-dependent C-T and T-T mismatches. Compared to ND- and naphthyridine-azaquinolone hybrid (NA)-immobilized sensor surfaces, with affinity to mismatches composed of purine nucleotide bases, the damND-immobilized surface was useful for the detection of the mismatches composed of pyrimidine nucleotide bases. 相似文献
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