Fig. 3.

RT-PCR validation of 21 predicted novel genes. a Amplification of a fragment of PfAlba3 (PF3D7_1006200) using genomic DNA (middle lane) or cDNA prepared from DNase-treated total RNA (right lane) as a template. Primers were designed on both sides of intron 1, yielding a 429 bp PCR product from genomic DNA and a 164 bp PCR product from cDNA. The presence of a single 164 bp PCR product amplified from cDNA confirms the absence of gDNA contamination. b Out of our 231 novel candidate genes, we chose 21 regions for validation using reverse transcription polymerase chain reaction (RT-PCR). The top panel shows amplification products using DNase-treated cDNA as a template, while the bottom panel shows the control reactions using genomic DNA as a template. Of the 21 gene tested, we were able to amplify 20 of the predicted regions. As a control, we were unable to amplify a fragment of intergenic region that was not predicted to contain any genes (marked as “intergenic”)