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Nels A Olson

Publications and source records attributed to Nels A Olson.

3 recordsLinked to original sources

DNA profiling by capillary array electrophoresis with non-covalent fluorescent labeling.

Increasing need for large-scale DNA profiling necessitated the development of automated electrophoresis based methods enabling rapid, high performance analysis of nucleic acids in a wide molecular-mass range. In this paper, we report on the adaptation of a commercial 96-capillary array electrophoresis (CAE) instrument for high-throughput DNA fragment analysis and the evaluation of the effects of different non-covalent DNA staining dyes on separation efficiency. The applicability of different color internal fluorescent standards is shown with mathematical spectral overlap correction algorithms. Large-scale quality control assessment of oligonucleotide probes using non-covalent fluorophore labeling is also demonstrated. The method requires small sample amounts, offers automation and quantification capabilities to accommodate modern biotechnology industry needs.

Algorithms↗

Large-scale carbohydrate analysis by capillary array electrophoresis: part 1. Separation and scale-up.

A 96-capillary array electrophoresis (CAE) instrument has been adapted for large-scale mono- and oligosaccharide analysis and characterization. Operational protocols and data processing tools have been developed to optimize the CAE system for this application. Effects of different additives to the running buffer on efficiency and capillary-to-capillary performance reproducibility have been studied.

Buffers↗

Large-scale carbohydrate analysis by capillary array electrophoresis: part 2. Data normalization and quantification.

Automated 96-capillary array electrophoresis (CAE) methodology described in the first part of the present work offered large-scale high-performance profiling of oligo- and monosaccharides to fulfill the needs of bioindustrial laboratories. Sensitivity at low nanomolar concentration, good resolving power and reliability achieved in the experiments is invaluable for monitoring reaction products from enzymatic polysaccharide digestion with numerous applications in agricultural, chemical and food industries. In addition to optimization of mono- and oligosaccharide separations in CAE system and necessary operational protocol modifications, capillary-to-capillary and run-to-run variation in migration time and signal intensity necessitated development of data normalization tools. Internal fluorescent standards have been incorporated into the analysis aiding migration time normalization and CAE trace alignment. Data processing, visualization, and programming tools have been developed along with quantification approaches.

Biotechnology↗