Sense and antisense transcription of gene with ID 76852. (A) IGV (Integrative Genome Viewer) output of the alignment of RNA-seq reads to the genome region corresponding to gene transcript ID 76852 under each condition. Blue reads represent sense RNAs, red reads represent antisense RNAs. (B) RT-PCR using gene specific primers (indicated by a yellow line) on oligo(dT) primed cDNA. The expected band size from spliced, sense transcripts is 476 bp and the size of the non-spliced antisense transcripts is 670 bp and has the same size as products from reactions run on gDNA. (C) Strand-specific RT-PCR using one of the standard PCR primers to synthesise cDNA from one strand only and then the PCR step was performed by using the same primer together with the opposing gene-specific primer. In 24 h straw and 5 h glucose both antisense (extend differently over both introns) and sense (differently spliced introns) transcripts are present, explaining the presence of multiple PCR products.