DNA separation by capillary electrophoresis with hydrophilic substituted celluloses as coating and sieving polymers. Application to the analysis of genetically modified meals.
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.
Giovannoli
Cristina
C
Department of Analytical Chemistry, Via P Giuria 5, University of Turin, 10125 Turin, Italy. cristina.giovannoli@unito.it
Anfossi
Laura
L
Tozzi
Cinzia
C
Giraudi
Gianfranco
G
Vanni
Adriano
A
D000327:Adsorption; D002021:Buffers; D002482:Cellulose / Q000031:analogs & derivatives*; D018744:DNA, Plant / Q000032:analysis* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary; D005504:Food Analysis; D030841:Food, Genetically Modified; D030821:Plants, Genetically Modified; D013025:Glycine max / Q000235:genetics; D003313:Zea mays / Q000235:genetics
D002021:Buffers; D018744:DNA, Plant; D002482:Cellulose