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|a eng
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|a Sano, M
|e verfasserin
|4 aut
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|a Electrophoretic mobility shift scanning using an automated infrared DNA sequencer
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|c 2001
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|a Text
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|a ohne Hilfsmittel zu benutzen
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|a Date Completed 18.04.2002
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a Electrophoretic mobility shift assay (EMSA) is widely used in the study of sequence-specific DNA-binding proteins, including transcription factors and mismatch binding proteins. We have established a non-radioisotope-based protocol for EMSA that features an automated DNA sequencer with an infrared fluorescent dye (IRDye) detection unit. Our modification of the elec- trophoresis unit, which includes cooling the gel plates with a reduced well-to-read length, has made it possible to detect shifted bands within 1 h. Further, we have developed a rapid ligation-based method for generating IRDye-labeled probes with an approximately 60% cost reduction. This method has the advantages of real-time scanning, stability of labeled probes, and better safety associated with nonradioactive methods of detection. Analysis of a promoter from an industrially important filamentous fungus, Aspergillus oryzae, in a prototype experiment revealed that the method we describe has potential for use in systematic scanning and identification of the functionally important elements to which cellular factors bind in a sequence-specific manner
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|a Ohyama, A
|e verfasserin
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|a Takase, K
|e verfasserin
|4 aut
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|a Yamamoto, M
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|a Machida, M
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|i Enthalten in
|t BioTechniques
|d 1993
|g 31(2001), 5 vom: 16. Nov., Seite 1056-8, 1060, 1062
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|x 0736-6205
|7 nnns
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| 773 |
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|g volume:31
|g year:2001
|g number:5
|g day:16
|g month:11
|g pages:1056-8, 1060, 1062
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