DNA fragments derived from various amplicons of the region spanning SP_1922 – SP_1926. RT-PCR on S. pneumoniae strain TIGR4 grown in rich medium (THYE) showed amplification products of the expected sizes. Lane 1 = Hyperladder I (Bioline). The remaining lanes represent pairs of primers tested on (i) unprocessed RNA (negative control, confirms absence of contaminating gDNA), cDNA (query), and gDNA (positive control), all from strain TIGR4. Primer pairs spanned: SP_1922 and SP_1923 (ply) (lanes 2–4 & 5–7), SP_1923 and SP_1924 (lanes 8–10), SP_1924 and SP_1926 (lanes 11–13), SP_1922 intragenic region (lanes 14–16), SP_1923 intragenic region (lanes 17–19), and SP_1926 and SP_1927 (negative control, lanes 20–22). It should be noted that the band of the highest molecular weight in the cDNA lane of the negative control (lane 21) is smaller than the band in the gDNA lane (22) and is part of a non-specific smear of amplification.