The G-less cassette is a reporter gene that encodes a transcript lacking guanine nucleotides in the sense strand of the DNA (hence "G-less"). A plasmid containing such a gene  is located downstream of a MCS. After the promoter is inserted into the MCS, transcription proceeds with the addition of radiolabeled UTP, CTP, and ATP (as well as non-radiolabeled/cold nucleotides) and continues until the end of the G-less cassette is reached and guanine residues are once again apparent in the sense strand of the DNA. The absence of GTP in vitro results in transcription being prematurely terminated at the first guanine residue in the sense strand following the cassette. Gel electrophoresis is performed on the transcription products and the amount of radioactivity is quantified by autoradiography or phosphorimaging to determine the strength of the promoter of interest.