Affinity capillary electrophoresis with magnetic beads for multiplex quantitative analysis of bacterial 16S rRNA(METHODS)
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概要
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We have developed a novel method for microbial community analysis of bacterial 16S rRNAs based on affinity capillary electrophoresis using 16S rRNA-conjugated magnetic beads. We called this method magnetic beads affinity capillary electrophoresis (MB-ACE) which can be used for sequential and quantitative analysis of 16S rRNA. In this method, RNA extracted from a microbial community is biotin-modified and mixed with streptavidin-modified paramagnetic beads. This mixture is then injected into a capillary and localized in the middle of the capillary using a magnet held adjacent to the capillary. Subsequently, a fluorescent-labeled probe to detect the target 16S rRNA is injected into the capillary, and voltage is applied. The probe trapped on the RNA is dissociated by formamide and detected at its anodic end by measuring the fluorescence. Next, another fluorescent probe is injected, and thus the target 16S rRNA in the sample is quantified one by one. MB-ACE was used for the quantification of the 16S rRNAs of Escherichia coli and Pseudomonas putida in samples that were prepared by mixing RNA extracted from activated sludge and 16S rRNAs prepared by in vitro transcription. The two types of 16S rRNAs were quantified, indicating that MB-ACE can be used for sequential quantitative analysis of bacterial 16S rRNAs.
- 社団法人日本生物工学会の論文
著者
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Tsuneda Satoshi
Department of Life Science and Medical Bioscience, Waseda University
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Torimura Masaki
Research Institute for Environmental Management Technology, National Institute of Advanced Industria
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SEKIGUCHI YUJI
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science
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NODA NAOHIRO
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science
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Tsuneda Satoshi
Department Of Chemical Engineering Waseda University
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Noda Naohiro
Institute For Biological Resources And Functions National Institute Of Advanced Industrial Science A
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Noda Naohiro
Institute For Biological Resources And Functions National Institute Of Advanced Industrial Science A
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Adachi Ken
Department of Chemical Engineering, Waseda University
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Yamaguchi Masahiro
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science
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Nakashige Makoto
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science
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Kanagawa Takahiro
Department of Bioenvironmental Design, Kyoto Gakuen University
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Torimura Masaki
Research Institute For Environmental Management Technology National Institute Of Advanced Industrial
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Adachi Ken
Department Of Chemical Engineering Waseda University:institute For Biological Resources And Function
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Sekiguchi Yuji
Institute For Biological Resources And Functions National Institute Of Advanced Industrial Science A
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Nakashige Makoto
Institute For Biological Resources And Functions National Institute Of Advanced Industrial Science A
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Kanagawa Takahiro
Department Of Bioenvironmental Design Kyoto Gakuen University
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Yamaguchi Masahiro
Institute For Biological Resources And Functions National Institute Of Advanced Industrial Science A
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