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 streptavidin-biotin system and subsequently used to monitor different unit operations in molecular biology, e.g., DNA strand separation, DNA hybridization kinetics, and enzymatic modifications. A model system comprising six oligonucleotides was used, which can be assembled into a 69-bp double-stranded DNA fragment. Using this system, the biosensor approach was employed to analyze multistep solid-phase gene assembly and the performance of different enzymes routinely used for the synthesis and manipulation of DNA. In addition, a concept for the determination of single-point mutations in DNA samples is described.
Nilsson
P
P
Department of Biochemistry and Biotechnology, Royal Institute of Technology (KTH), Stockholm, Sweden.
Persson
B
B
Uhlén
M
M
Nygren
P A
PA
D001483:Base Sequence; D015374:Biosensing Techniques; D004247:DNA / Q000032:analysis* / Q000096:biosynthesis; D004720:Endonucleases / Q000494:pharmacology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA / Q000379:methods
D004247:DNA; D004720:Endonucleases