View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Digital.CSIC 1 TITLE: Development of a Real-Time PCR method for the identification of 2 Atlantic mackerel (Scomber scombrus) 3 4 AUTHORS: Amaya Velasco1*, Ana Sánchez1, Icíar Martínez2, Francisco J. 5 Santaclara1, Ricardo I. Pérez-Martín1, Carmen G. Sotelo1 6 1Instituto de Investigaciones Marinas (IIM-CSIC), Eduardo Cabello 6, E-36208 7 Vigo, Spain. 8 2Plentzia Marine Station; University of the Basque Country &IKERBASQUE, 9 Basque Foundation for Science, Areatza, E-48620 Plentzia, Spain. 10 11 *Corresponding author: Amaya Velasco, Instituto de Investigaciones Marinas 12 (IIM-CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain. Tel: +34-986231930, Fax: 13 +34-986292762, Email:
[email protected] 14 ABSTRACT: A Real Time-PCR method based on TaqMan technology for the 15 identification of Scomber Scombrus has been developed. A system of specific 16 primers and a Minor Groove Binding (MGB) TaqMan probe based on 17 sequences of the mitochondrial cytochrome b region was designed. The 18 method was successfully tested in 81 specimens of Scomber scombrus and 19 related species and validated in 26 different commercial samples. An average 20 Threshold cycle (Ct) value of 15.3 was obtained with Scomber scombrus DNA. 21 With the other species tested fluorescence signal was not detected or Ct was 22 significantly higher (P<0.001). The efficiency of the assay was estimated to be 23 92.41%, with 100% specificity, and no cross reactivity was detected with any 24 other species. These results reveal that the developed method is a rapid and 25 efficient tool to unequivocally identify S.