Detection of genetically modified soybean using peptide nucleic acids (PNAs) and microarray technology.
Peptide nucleic acid (PNA) microarrays for the detection of Roundup Ready soybeans in food have been prepared. PNA probes are known to be more efficient and selective in binding DNA sequences than the analogous oligonucleotides and are very suitable to be used for diagnostics in food. PNAs of different lengths were carefully designed and synthesized by solid-phase synthesis on an automatic synthesizer adopting the BOC strategy. PNAs were purified by HPLC and characterized by HPLC/MS. The probes were spotted on a functionalized surface to produce a microarray to be hybridized with PCR products. DNA extracted from reference material was amplified using Cy3- and Cy5-labeled primers, and the fluorescent PCR products obtained were hybridized on the microarray. Two protocols were adopted: the hybridization with dsDNA or with ssDNA obtained by digestion with the enzyme lambda exonuclease. The best results were obtained using a 15-mer PNA probe in combination with the ssPCR product derived from enzymatic digestion. The method was applied to the analysis of a sample of certified transgenic soybean flour.
Germini
Andrea
A
Dipartimento di Chimica Organica e Industriale, Università degli Studi di Parma, Parco Area delle Scienze 17/A, 43100 Parma, Italy.
Mezzelani
Alessandra
A
Lesignoli
Francesca
F
Corradini
Roberto
R
Marchelli
Rosangela
R
Bordoni
Roberta
R
Consolandi
Clarissa
C
De Bellis
Gianluca
G
D018744:DNA, Plant / Q000032:analysis; D020411:Oligonucleotide Array Sequence Analysis; D020135:Peptide Nucleic Acids / Q000032:analysis*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013025:Glycine max / Q000235:genetics*
D018744:DNA, Plant; D020135:Peptide Nucleic Acids