REAL-TIME MONITORING OF DNA MANIPULATIONS USING BIOSENSOR TECHNOLOGY

The potential of real-time biospecific interaction analysis technology for applications in molecular biology is described. DNA fragments are immobilized onto a biosensor surface using the high-affinity streptav idin-biotin system and subsequently used to monitor different unit ope rations in molecular biology, e.g., DNA strand separation, DNA hybridi zation kinetics, and enzymatic modifications. A model system comprisin g six oligonucleotides was used, which can be assembled into a 69-bp d ouble-stranded DNA fragment. Using this system, the biosensor approach was employed to analyze multistep solid-phase gene assembly and the p erformance of different enzymes routinely used for the synthesis and m anipulation of DNA. In addition, a concept for the determination of si ngle-point mutations in DNA samples is described. (C) 1995 Academic Pr ess, Inc.