The DNA replication kit is suitable to kick start your single-molecule experiments to study the kinetics of enzyme activities involved in DNA replication. The biotinylated pKYB1 DNA containing a 25-nt 5′-overhang serves to load DNA polymerase enzymes (e.g., T7 DNA polymerase) to investigate the polymerization dynamics, proofreading activity, and pause states of DNA replication in real time.
Overall, the DNA replication kit will provide you with a ready-to-go system for single-molecule applications to:
- Obtain kinetics and mechanistic data of the different phases of the DNA replication process at the single-molecule level with millisecond and base-pair resolution.
- Correlate different tensions applied to a DNA – mimicking nucleotide miss-incorporation – with polymerization rates, exonucleolytic activity, and pause states of DNA polymerase enzymes.
- Test polymerase activity with the u-Flux™ Laminar Flow Microfluidics that, through its multiple channels, facilitates switching between different experimental conditions.
A schematic illustration depicting the exonucleolytic activity of T7 DNA polymerase as it removes single nucleotides from the substrate DNA molecule. The enzymatic activity can be measured with a base-pair resolution. Both ends of the DNA molecule are tethered to its respective optically trapped bead. Read more here: https://lumicks.com/application/dna-replication-optical-tweezers-fluorescence-microscopy/
- T7 DNA polymerase
- Biotinylated ds pKYB1 DNA with a 25-nt 5′-end overhang
- dNTP mix
- Streptavidin coated polystyrene beads (1.76 µm)
- Polymerase buffer
- Estimated for 2 weeks of measurements
- Estimated 1-3 weeks delivery time