Strand-Displacement Polymerization by Reverse Transcriptase Investigated using LUMICKS’ u-Flux™

The journal Nucleic Acids Research has recently published the paper “Pausing kinetics dominates strand-displacement polymerization by reverse transcriptase”. We would like to congratulate the authors!

Malik et al. (2017) used optical tweezers and, among other technologies, microfluidics to study the enzymatic activity of the murine leukemia Virus reverse transcriptase (RT). RT catalyzes the formation of an integration-competent double-stranded DNA from viral RNA and has the ability to displace a non-template strand simultaneously with DNA polymerization. The researchers initiated polymerization by first trapping the DNA using optical tweezers, and thereafter moving the tethered DNA to a channel containing activity buffer using LUMICKS’ u-Flux™ microfluidics laminar flow cell. The study shows that strand-displacement polymerization is frequently interrupted by the existence of a structured template ahead of the polymerization site and reveals important insights on the interactions between the RT and the template’s secondary structure.

The u-Flux™ provides multiple adjacent laminar flow channels that do not mix. No physical barriers are separating the laminar flows, which have been optimized for the most sensitive single-molecule experiments. Are you interested in using the instrument for your own research? Please feel free to contact us for a demo or quote.