New method of DNA isolation from two food additives suitable for authentication in polymerase chain reaction assays.
Locust bean gum and guar gum are galactomannans used as additives (E 410 and E 412, respectively) in the food industry as stabilizing agents. Analytical discrimination between the two additives in gums and foods is now feasible by molecular techniques. However, only complex and time-consuming DNA isolation protocols are available to date. We have developed simple improved protocols to obtain enough DNA suitable for PCR amplification from a few milligrams of commercial E 410 and E 412 additives (containing more than 75% polysaccharides). The suspension of additives in water or 10 mM Tris-HCl, pH 8.5, efficiently recovers DNA suitable for authentication in PCR assays. However, the Tris method was much more efficient for the extraction of DNA from E 410 than for E 412 additives. Conversely, the water method was the most suitable for detecting DNA extracted from E 412 or from E 410/E 412 mixtures. Combined with the use of the two specific ribosomal primer pairs previously designed, our methods are well-suited for a fast and simple high-throughput sample treatment of commercial gums for molecular certification.
Urdiaín
Mercedes
M
Microbiologia, Departament de Biología, Universitat de les Illes Balears, E-07071 Palma de Mallorca, Spain.
Doménech-Sánchez
Antonio
A
Albertí
Sebastián
S
Benedí
V Javier
VJ
Rosselló
Josep A
JA
D004247:DNA / Q000302:isolation & purification*; D005503:Food Additives / Q000032:analysis* / Q000737:chemistry*; D005685:Galactans / Q000032:analysis / Q000235:genetics; D008351:Mannans / Q000032:analysis / Q000235:genetics; D053149:Plant Gums; D016133:Polymerase Chain Reaction; D011134:Polysaccharides / Q000032:analysis / Q000235:genetics
D005503:Food Additives; D005685:Galactans; D008351:Mannans; D053149:Plant Gums; D011134:Polysaccharides; D004247:DNA; C007894:guar gum; C017471:locust bean gum