Additional file 2.
Cross-amplification controls for LWS primer specificity. (A) Locus-specific primer (LSP) sets, detailed in the methods, for each LWS (A180, P180, S180, and S180r) were individually used in an attempt to amplify off of LWS-containing plasmids (1.0E07 copies/reaction) under the RT-qPCR conditions detailed in the methods. Although cloned opsin gene (pGEM::LWS) sequences are not full length (i.e., incomplete open reading frame, ORF) their respective primer site locations are shared (although different in sequence) among all four genes. Plasmid inserts were generated using primers shown in part C. Amplification and dissociation curves are shown with corresponding agarose gel-electrophoresis images of resultant amplicons. 1kb+ DNA ladder was used (Invitrogen®). All reactions were run in parallel. Note the gel image corresponding to the S180r primer panel is not the same dimensions as the others, as lane gaps were not used during gel loading. (B) NAC and NTC controls are shown. (C) Table of Oligonucleotide primers and sequences used to generate LWS plasmids for cross-amplification experiments.
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Laver and Taylor BMC Evolutionary Biology 2011 11:81 doi:10.1186/1471-2148-11-81