The detection of oligonucleotide N3′ → P5′ phosphoramidate/RNA duplexes with capillary gel electrophoresis

1998 ◽  
Vol 19 (8-9) ◽  
pp. 1265-1269 ◽  
Author(s):  
Lawrence Dedionisio ◽  
Annette M. Raible ◽  
Sergei M. Gryaznov
Keyword(s):  
Gel Electrophoresis ◽  
Capillary Gel Electrophoresis ◽  
Rna Duplexes

scholarly journals Rapid capillary gel electrophoresis analysis of human milk oligosaccharides for food additive manufacturing in-process control

2021 ◽  
Author(s):  
Marton Szigeti ◽  
Agnes Meszaros-Matwiejuk ◽  
Dora Molnar-Gabor ◽  
Andras Guttman
Keyword(s):  
Additive Manufacturing ◽  
Process Control ◽  
Human Milk ◽  
Gel Electrophoresis ◽  
Process Analysis ◽  
Food Additive ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Capillary Gel Electrophoresis

AbstractIndustrial production of human milk oligosaccharides (HMOs) represents a recently growing interest since they serve as key ingredients in baby formulas and are also utilized as dietary supplements for all age groups. Despite their short oligosaccharide chain lengths, HMO analysis is challenging due to extensive positional and linkage variations. Capillary gel electrophoresis primarily separates analyte molecules based on their hydrodynamic volume to charge ratios, thus, offers excellent resolution for most of such otherwise difficult-to-separate isomers. In this work, two commercially available gel compositions were evaluated on the analysis of a mixture of ten synthetic HMOs. The relevant respective separation matrices were then applied to selected analytical in-process control examples. The conventionally used carbohydrate separation matrix was applied for the in-process analysis of bacteria-mediated production of 3-fucosyllactose, lacto-N-tetraose, and lacto-N-neotetraose. The other example showed the suitability of the method for the in vivo in-process control of a shake flask and fermentation approach of 2′-fucosyllactose production. In this latter instance, borate complexation was utilized to efficiently separate the 2′- and 3-fucosylated lactose positional isomers. In all instances, the analysis of the HMOs of interest required only a couple of minutes with high resolution and excellent migration time and peak area reproducibility (average RSD 0.26% and 3.56%, respectively), features representing high importance in food additive manufacturing in-process control. Graphical abstract

Optimized Large Scale Synthesis of Phosphorothioate Oligonucleotides on PS-PEG Supports

1997 ◽  
Vol 52 (1) ◽  
pp. 110-116
Author(s):  
Michael Gerster ◽  
Martin Maier ◽  
Nils Clausen ◽  
Jens Schewitz ◽  
Ernst Bayer
Keyword(s):  
Gel Electrophoresis ◽  
Continuous Flow ◽  
Large Scale ◽  
Electrospray Mass Spectrometry ◽  
Small Scale ◽  
Batch Mode ◽  
Crucial Step ◽  
Capillary Gel Electrophoresis ◽  
Phosphoramidite Chemistry ◽  
Large Scale Synthesis

Sulphurization is a crucial step during synthesis of phosphorothioate oligonucleotides. Insufficient reaction leads to inhomogeneous products with phosphodiester defects and subsequently to destabilization of the oligomers in biological media. To achieve a maximum extent of sulphur incorporation, various sulphurizing agents have been investigated. Solely, the use of Beaucage reagent provided satisfactory results on PS-PEG supports. Based on our investigations in small scale synthesis (1 μmol) with continuous-flow technique, upscaling to the 0.1-0.25 mmolar range has been achieved using a peptide synthesizer. The syntheses were performed in batch mode with standard phosphoramidite chemistry. Additionally, large scale synthesis of a phosphodiester oligonucleotide has been carried out on PS-PEG with optimized protocols and compared to small scale synthesis on different supports. Products were analysed by 31P NMR, capillary gel electrophoresis and electrospray mass spectrometry. An extent of sulphurization of 99% and coupling effiencies of more than 99% were obtained and the products proved to have similar purity compared to small scale syntheses on CPG

Capillary gel electrophoresis of nucleic acids in pluronic F127 copolymer liquid crystals

1999 ◽  
Vol 49 (S1) ◽  
pp. S65-S71 ◽  
Author(s):  
R. L. Rill ◽  
Y. Liu ◽  
B. A. Ramey ◽  
D. H. Van Winkle ◽  
B. R. Locke
Keyword(s):  
Liquid Crystals ◽  
Nucleic Acids ◽  
Gel Electrophoresis ◽  
Pluronic F127 ◽  
Capillary Gel Electrophoresis ◽  
Pluronic F127 Copolymer
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