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Selection of in silico SNP discovery panel (Additional file 2). cDNA library development (Table 1, Additional file 3). Sequence assembly (MIRA 3.1) (Table 2). cDNA library sequencing (Roche GS FLX 454). Contig alignment (Roche gsMapper) (Additional file 4). SNP candidate selection
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Selection of in silico SNP discovery panel (Additional file 2) cDNA library development (Table 1, Additional file 3) Sequence assembly (MIRA 3.1) (Table 2) cDNA library sequencing (Roche GS FLX 454) Contig alignment (Roche gsMapper) (Additional file 4) SNP candidate selection (Additional file 5) > one polymorphic base/locus Base call of ‘N’ or ‘– ’ Frequency <100% Sequencing depth <5 Optimized SNP candidates Primer design (BatchPrimer3) SNP Validation (High-resolution melt analysis) (Figure 2)