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. 2009 Feb 6;5(2):e1000364. doi: 10.1371/journal.pgen.1000364

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Gaillard et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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In vitro transcription assay of WCEs from BY4741 (wt), med15, bur2, fun12, bre1, spt20, tpd3, and not5 strains. Each reaction was stopped after 30 min, treated with RNaseT1 and run in a 6% PAGE. Two bands from each G-less cassette were obtained, probably due to incomplete action of RNaseT1. Efficiency of transcription elongation was determined as the percentage of transcripts that reach the 376-nt cassette in respect to those that cover the 84-nt cassette. Radioactivity incorporated into the cassettes was quantified in a Fuji FLA3000 and normalized with respect to the C content of each cassette. The mean value of the wt (84%) was normalized to 100%. Mean value and standard deviation of three independent experiments are shown.