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. 2004 Apr;72(4):1874–1884. doi: 10.1128/IAI.72.4.1874-1884.2004

FIG. 5.

FIG. 5.

Multiplex RT-PCR analysis of H. ducreyi lspB-containing transcripts. The following templates were included in the reaction mixtures: lane 1, no template (negative control); lanes 2 and 4, 100 ng of H. ducreyi wild-type strain 35000 genomic DNA (positive control); lanes 3 and 5, 100 ng of H. ducreyi lspA1 mutant genomic DNA (positive control); lanes 6, 8, 10, and 12, 1 μg of H. ducreyi wild-type strain 35000 total RNA; lanes 7, 9, 11, and 13, 1 μg of H. ducreyi lspA1 mutant total RNA. The primer sets included in the reaction mixtures were as follows: lanes 1 to 3, 6, 7, 10, and 11 contained both pal-specific (354-bp product) and lspB-lspA2 (431-bp product) primers; lanes 4, 5, 8, 9, 12, and 13 contained both pal-specific (354-bp product) and lspB-specific (226-bp product) primers. The reaction mixtures loaded in lanes 10 to 13 were not subjected to the RT step of the RT-PCR procedure and served as controls to detect DNA contamination of RNA preparations. Lane M contained DNA size markers.