In this chapter, we present methods for denaturing and native gel electrophoresis of nucleic acids based on fluorescently labeled probes and automatic signal detection by a deoxyribonucleic acid sequencer. Specific examples are given for the determination of ribonucleic acid (RNA) fragmentation patterns, of (deoxy)ribozyme kinetics, and of direct or competitive gel-shift assays. These methods can replace widely used radioisotope-based protocols, for example, for secondary structure mapping of RNA and for the characterization of nucleic acid ligand interactions.
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http://dx.doi.org/10.1385/1-59259-823-4:261 | DOI Listing |
Biotechnol Prog
March 2019
National Research Council Canada, Montreal, QC, H4P 2R2, Canada.
This study describes the application of the multivariate curve resolution (MCR) analysis technique for real-time analysis of culture fluorescence during recombinant Pichia pastoris cultivation in a bioreactor. Fluorescence spectra were acquired with an on-line dual excitation wavelength fluorometer and then used to develop a real time MCR-based bioprocess monitoring and diagnostics tool. Initial bioreactor experiments using two similar recombinant antibody secreting P.
View Article and Find Full Text PDFFood Addit Contam Part A Chem Anal Control Expo Risk Assess
May 2018
a Department of Physical and Analytical Chemistry, Faculty of Experimental Sciences , University of Jaén, Jaén , Spain.
One of the most used agrochemicals in agricultural production, nitenpyram (NTP), has been determined by using a flow-through optosensing device based on Photochemically Induced Fluorescence detection. The combination of both methodologies allows, on one hand, a quick on-line photodegradation of NTP and, on the other hand, the preconcentration, quantification and desorption of the fluorescent photoproduct generated when retained on Sephadex QAE-A25 as solid support, which was monitored at 295 and 362 nm for excitation and emission, respectively. The proposed analytical method presents a detection limit of 500 pg mL by using Multicommutated Flow Injection Analysis.
View Article and Find Full Text PDFSensors (Basel)
January 2017
Department of Energy Technology, Aalborg University, Esbjerg 6700, Denmark.
This article is a feasibility study on using fluorescence-based oil-in-water (OiW) monitors for on-line dynamic efficiency measurement of a deoiling hydrocyclone. Dynamic measurements are crucial in the design and validation of dynamic models of the hydrocyclones, and to our knowledge, no dynamic OiW analysis of hydrocyclones has been carried out. Previous studies have extensively studied the steady state efficiency perspective of hydrocyclones, and have related them to different key parameters, such as the pressure drop ratio (PDR), inlet flow rate, and the flow-spilt.
View Article and Find Full Text PDFSensors (Basel)
December 2012
Department of Engineering Enzo Ferrari, University of Modena and Reggio Emilia,via Vignolese 905, Modena I-41125, Italy.
The design, development and performance evaluation of a fluorescence-based pH sensor for on-line measurements is presented. The pKa of the sensing element has been calculated to be 7.9, thus the sensor is suitable for measurement of near neutral solutions.
View Article and Find Full Text PDFLab Chip
November 2012
Translational Centre for Regenerative Medicine, University of Leipzig, Germany.
Impedance flow cytometry (IFC) was evaluated as a possible alternative to fluorescence-based methods for on-line quality monitoring of hybridoma cells. Hybridoma cells were cultured at different cell densities and viability was estimated by means of IFC and fluorescence-based flow cytometry (FCM). Cell death was determined by measuring the impedance phase value at high frequency in low conductivity buffer.
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