Validating MS data using RT-PCR. RNA fragments of the expected sizes were observed, indicating that these un-annotated genes are transcribed. Figure shows the RT-PCR verification results for six novel genes. Amplified PCR products were electrophoresed on a 2.5% agarose gel and visualized by ethidium bromide staining. BIO07235, BIO01608, BIO43803, and BIO50043 were amplified and loaded in lanes 1-4, respectively; a negative control (noncoding DNA sequence) was loaded in lane 5 (cDNA as template) and lane 6 (genomic DNA as template); Lane 7, positive control (housekeeping gene, ipaD); Lane M, GeneRuler™ 50 bp DNA Ladder (Fermentas GmbH, Germany).