Development of sensitive crop-specific polymerase chain reaction assays using 5S DNA: applications in food traceability.
The 5S intergenic spacers were amplified using a common pair of primers and sequenced from four species (Brassica napus, Zea mays, Helianthus annuus, and Glycine max). Crop-specific assays were developed from primers designed from the spacers and tested to amplify corresponding DNAs in both conventional end-point and real-time polymerase chain reactions (PCRs). The high copy numbers of the 5S DNA in plants make it possible to detect very small amounts of DNA using this marker. This sensitivity made it possible to compare different DNA extraction methods for highly processed food products using 5S spacers, even allowing dilution of templates to overcome PCR inhibition.
Doveri
Silvia
S
Crop Genetics Group, NIAB, Huntingdon Road, Cambridge CB3 0LE, United Kingdom.
Lee
David
D
D017931:DNA Primers; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005504:Food Analysis / Q000379:methods*; D007438:Introns; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018744:DNA, Plant