Circular dichroism spectroscopy of DNA duplexes at near-biological concentrations

Circular dichroism (CD) of nucleic acids has been typically carried out at sample concentrations below 10 μM, which is far lower than nucleic acid concentrations in biological systems. Attempts to study nucleic acid conformations by CD at higher concentrations using 10 and 1 mm pathlength cuvettes led to instrument artifacts. By shortening the light pathlength to around 0.1 mm, we herein report the first CD profiles of nucleic acids at sub-mM concentrations, which are relevant to nucleic acid concentrations in cellular cytoplasm and nucleus. These CD experimental conditions will allow future conformational studies of nucleic acids under biologically relevant conditions.