Secondary verification of RNA-seq results by RT-PCR. RT-PCR verifies that lecA and leu1 are anaerobically induced, and that NGO0108, lldD, trx1, hemO, and nqrA are anaerobically repressed. RT-PCR of the murA transcript (NGO1981), which was shown to have an equal expression level aerobically and anaerobically (See additional file 1: Supplementary Table S1), was used as a loading control. (-RT) signifies that reverse transcriptase was not included in the reaction. For the anaerobically induced genes lexA/erpA and NG1428/dnaK, primers were used that amplified across the 3'end of the lexA coding region into the 5' end of the erpA coding region, and from the 3' end of the NG1428 coding region into 5'end of the dnaK coding region, respectively. Amplification across these intergenic regions suggests that these genes are cotranscribed.
Isabella and Clark BMC Genomics 2011 12:51 doi:10.1186/1471-2164-12-51