Figure 1.
The universal primer system: Scheme of base-specific cleavage and MALDI-TOF MS based resequencing. PCR amplification of the DNA region of interest is followed by T7-mediated in vitro transcription and base-specific cleavage. RNA transcripts of the target regions are cleaved in four reactions corresponding to virtually each of the four bases. This corresponds to an RNAse-specific cleavage at U and C of the forward as well as of the reverse RNA transcript. MALDI-TOF spectra of all four cleavage reactions are subject to subsequent mutation analysis.
