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Translation toeprinting assays using fluorescently labeled primers and capillary electrophoresis
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UNSPECIFIED (2005) Translation toeprinting assays using fluorescently labeled primers and capillary electrophoresis. BIOTECHNIQUES, 38 (3). pp. 397-400. ISSN 0736-6205.
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Abstract
The protocol described here is an adapted version of the toeprinting assay in which the oligonucleotide used to prime the reverse transcription step is labeled with a fluorescent dye instead of P-32. By using a fluorescent dye, the results of the assay arc obtained within one hour by direct electrophoresis of the samples on an automated sequencing machine. This eliminates the need to cast and run polyacrylamide gels and to wait to expose dried gels. We show that an identical toeprint was found for the chloramphenicol acetyltransferase transcript using this nonradioactive Method, which is in agreement with the previously published P-32-labeled method. Furthermore, in addition to being a faster and safer method, a larger region of sequence can be analyzed with one primer in a single experiment.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Journal or Publication Title: | BIOTECHNIQUES | ||||
Publisher: | EATON PUBLISHING CO | ||||
ISSN: | 0736-6205 | ||||
Official Date: | March 2005 | ||||
Dates: |
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Volume: | 38 | ||||
Number: | 3 | ||||
Number of Pages: | 4 | ||||
Page Range: | pp. 397-400 | ||||
Publication Status: | Published |
Data sourced from Thomson Reuters' Web of Knowledge
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