I read with interest the article “Widespread Antisense Transcription in Escherichia coli” by Dornenburg and colleagues (1) for the new journal mBio. However, I was surprised by the lack of documentation of the methods used. The description is insufficient to allow attempted replication of the results. There is no mention of how trace DNA might have been excluded from the protocol.Furthermore, the discussion does nothing to address possible alternative interpretations of the result and what controls were used.I think that, given the large numbers of transcripts found (about a quarter of the genome), controls and interpretation should have been a significant part of the paper. These are issues that reviewers generally expect addressed before publication.