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DNA separation by capillary electrophoresis with hydrophilic substituted celluloses as coating and sieving polymers. Application to the analysis of genetically modified meals
Author(s) -
Giovannoli Cristina,
Anfossi Laura,
Tozzi Cinzia,
Giraudi Gianfranco,
Vanni Adriano
Publication year - 2004
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200401797
Subject(s) - hydroxyethyl cellulose , chromatography , capillary electrophoresis , coating , ethidium bromide , polymer , chemistry , cellulose , capillary action , reproducibility , electrophoresis , adsorption , tris , dna , materials science , organic chemistry , biochemistry , composite material
A coating procedure based on the physical adsorption of hydroxypropyl cellulose onto the wall of a capillary column has been successfully used for the separation of DNA fragments up to 500 bp. The method uses a running Tris‐phosphate‐EDTA buffer containing 2‐hydroxyethyl cellulose as sieving polymer. The separation procedure shows good reproducibility (measured as RSD%) for consecutive runs (< 0.64), for different days (< 1.15) and capillaries (< 2.15), short analysis times, and a long coating lifetime. Good reproducibility and efficiency are even achieved by performing the separation in the presence of additives such as ethidium bromide and mannitol. The method is applied to the detection of GMOs in soybean and maize meals with an accurate evaluation of the length of DNA sequences, previously amplified by polymerase chain reaction.