Consolandi Clarissa, Palmieri Luisa, Doveri Silvia, Maestri Elena, Marmiroli Nelson, Reale Silvia, Lee David, Baldoni Luciana, Tosti Nicola, Severgnini Marco, De Bellis Gianluca, Castiglioni Bianca
Institute of Biomedical Technologies, National Research Council, LITA, Via Fratelli Cervi 93, I-20090 Segrate (MI), Italy.
J Biotechnol. 2007 May 1;129(3):565-74. doi: 10.1016/j.jbiotec.2007.01.025. Epub 2007 Feb 9.
Molecular methodologies are increasingly being developed to assess the origin and authenticity of raw organic materials and processed food products. Here we describe the application of a microarray-based assay for single nucleotide polymorphisms (SNPs) identification in olive cultivars. The assay distinguishes alleles in a ligation detection reaction (LDR), with subsequent fluorescent detection by hybridization on a universal array (UA). The LDR-UA approach was used to detect 17 SNPs in olive genomic sequences previously amplified by PCR from fresh olive leaves. Genotype calls obtained with the LDR-UA were in full agreement with those determined by direct sequencing. The panel of 17 SNPs is sufficient to discriminate 49 olive varieties selected among the most widely cultivated for olive oil production in the Mediterranean area.
越来越多的分子方法被开发出来,用于评估生有机材料和加工食品的来源及真实性。在此,我们描述了一种基于微阵列的检测方法在橄榄品种单核苷酸多态性(SNP)鉴定中的应用。该检测方法在连接检测反应(LDR)中区分等位基因,随后通过在通用阵列(UA)上杂交进行荧光检测。LDR-UA方法用于检测先前从新鲜橄榄叶中通过PCR扩增的橄榄基因组序列中的17个SNP。用LDR-UA获得的基因型判定结果与直接测序确定的结果完全一致。这17个SNP足以区分从地中海地区最广泛种植用于生产橄榄油的品种中选出的49个橄榄品种。