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1.
Anal Bioanal Chem ; 407(5): 1499-503, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25433688

ABSTRACT

A selective separation method using a poly(methylmethacrylate) microchip was developed for 7-amino-4-methylcoumarin-labeled saccharides in a crude reaction mixture. In an alkaline borate buffer, saccharide derivatives formed strong anionic borate complexes. These complexes moved from the cathode to the anode in an electric field and were detected near the anode. Excess labeling reagents and other foreign substances remained at the inlet reservoir. A confocal fluorimetric detection system enabled the determination of monosaccharide derivatives with good linearity between at least 5 and 100 nM, corresponding to 50 fmol to 1 pmol per injection. The lower limit of detection (signal-to-noise = 5) was 2 nM. The sensitivity and linear quantitation range were comparable to reported values using fluorometric detection, capillary electrophoresis, or liquid chromatography. Application of the method showed excellent resolution in the analysis of O-linked glycans chemically released from glycoproteins.


Subject(s)
Electrophoresis, Microchip/methods , Oligosaccharides/chemistry , Oligosaccharides/isolation & purification , Coumarins/chemistry , Electrophoresis, Microchip/instrumentation , Glycoproteins/chemistry , Sensitivity and Specificity
2.
Nucleic Acids Res ; 41(Database issue): D732-7, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23172286

ABSTRACT

During animal development, cells undergo dynamic changes in position and gene expression. A collection of quantitative information about morphological dynamics under a wide variety of gene perturbations would provide a rich resource for understanding the molecular mechanisms of development. Here, we created a database, the Worm Developmental Dynamics Database (http://so.qbic.riken.jp/wddd/), which stores a collection of quantitative information about cell division dynamics in early Caenorhabditis elegans embryos with single genes silenced by RNA-mediated interference. The information contains the three-dimensional coordinate values of the outlines of nuclear regions and the dynamics of the outlines over time. The database provides free access to 50 sets of quantitative data for wild-type embryos and 136 sets of quantitative data for RNA-mediated interference embryos corresponding to 72 of the 97 essential embryonic genes on chromosome III. The database also provides sets of four-dimensional differential interference contrast microscopy images on which the quantitative data were based. The database will provide a novel opportunity for the development of computational methods to obtain fresh insights into the mechanisms of development. The quantitative information and microscopy images can be synchronously viewed through a web browser, which is designed for easy access by experimental biologists.


Subject(s)
Caenorhabditis elegans/embryology , Caenorhabditis elegans/genetics , Databases, Genetic , Animals , Cell Division/genetics , Genes, Helminth , Internet , RNA Interference
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