text: The G-less assay can be performed on a circular plasmid to measure levels of transcription. A circular plasmid provides a more efficient template in many systems when compared to other assays such as runoff transcription, in which a cleaved end is required. This method generates radiolabeled transcripts very efficiently because it bypasses the unnecessary process of performing other indirect mRNA product measurements. The promoter is inserted into a circular plasmid containing the G-less cassette, which will generate a transcript of a certain length that omits random and nonspecific transcription throughout the plasmid. Most crude systems, such as HeLa nuclear extracts, are used because they contain low amounts of contaminating GTP that lead to background transcription and may occasionally cause random transcription to read through the G-less cassette.