Evaluation of purification procedures of DNA from maize-meal samples by exploiting different analytical techniques for the assessment of DNA quality.
Two different approaches generally applied to achieve purification of DNA extracted from cells were compared: precipitation by organic solvents and enzymatic treatments. We investigated various experimental protocols reported in literature by evaluating DNA purity, integrity and yield. Reliability of analytical techniques normally employed to assess DNA purity and quantity was studied and comments and conclusions were suggested by comparing results obtained by different analytical techniques. Enzymatic treatments prove to be unable of increasing DNA purity while determining a significant degradation. In contrast, optimised conditions for solvent precipitation enabled a sharp increase of DNA purity to be obtained, associated with the maintenance of the initial DNA integrity. The application of the optimised protocol to maize-meal samples allowed us to achieve a good PCR amplification even with those samples which gave poor amplification by following the protocol recommended by the Italian legislation in force for GMO detection in food.
Vanni
Adriano
A
Department of Analytical Chemistry, University of Turin, Via Giuria 5, 10125 Torino, Italy. adriano.vanni@unito.it
Anfossi
Laura
L
Giovannoli
Cristina
C
Oddenino
Leila
L
Giraudi
Gianfranco
G
D011232:Chemical Precipitation; D002623:Chemistry Techniques, Analytical / Q000379:methods; D004247:DNA / Q000032:analysis*; D005502:Food / Q000592:standards; D007558:Italy; D030781:Organisms, Genetically Modified; D016133:Polymerase Chain Reaction; D012997:Solvents; D003313:Zea mays / Q000737:chemistry / Q000592:standards*
D012997:Solvents; D004247:DNA