New Genetic Tools for Use in the Rhizobiaceae and Other Bacteria

1 0 000copies of plasmid DNA described in this paper.Indeed, the accessibility of the target is higher on plasmid DNA than on genomic DNA that has a more complex conformational organization.In addition, replicate intermediates of the plasmid are single-stranded and so are a template for NASBA amplification.Therefore, NASBA is not a suitable method to detect transcription of plasmid DNA.From these results, we determined that for the highly sensitive detection of eukaryotic gene transcription with NASBA, massive contamination of the RNA sample with genomic DNA may be a problem if the template is multiple copy-number genes (e.g., HERV) and if the sample is derived from a tissue with high mitotic activity.In general, DNA molecules are not static replication and transcription events that lead to the formation of single-stranded regions or permit specific primer hybridization or amplification by NASBA.