Radioactive reverse transcription PCR (RT-PCR) analysis of strain SSB318 complemented with wt or C238/C239

Abstract

<p><b>Copyright information:</b></p><p>Taken from " and investigation of bacterial type B RNase P interaction with tRNA 3′-CCA"</p><p></p><p>Nucleic Acids Research 2007;35(6):2060-2073.</p><p>Published online 13 Mar 2007</p><p>PMCID:PMC1874595.</p><p>© 2007 The Author(s)</p> PCR products were analyzed on a 10% polyacrylamide/8 M urea gel. Lanes 1–30: total RNA from SSB318 complemented with wt (lanes 1–4 and 13-16), C238 (lanes 5–8, 17–20 and 25–30) or C239 (lanes 9–12 and 21–24) grown at 37°C in the absence of IPTG and in the presence of 2% xylose (w/v); amounts of total RNA were 200 ng in lanes 1–24, 26 and 29, 100 ng in lanes 25 and 28, and 400 ng in lanes 27 and 30. P : presence (+) or absence (−) of a xylose-inducible plasmid-encoded gene. Lanes 1–12 and 25–27: primers specific for ; lanes 13–24 and 28–30: primers specific for the mRNA encoding ribosomal protein S18 (S18). AMV: presence (+) or absence (−) of reverse transcriptase. For details on RT-PCR, see the Material and Methods section. Lanes 25–30 document that the amount of RT-PCR product was sensitive to RNA template concentration. The figure illustrates a representative experiment, but the results shown here were reproduced in five individual experiments using three independent total RNA preparations