Conformational
changes of single-stranded DNA (ssDNA) play an important
role in a DNA strand’s ability to bind to target ligands. A
variety of factors can influence conformation, including temperature,
ionic strength, pH, buffer cation valency, strand length, and sequence.
To better understand the effects of these factors on immobilized DNA
structures, we employ temperature-controlled electrochemical microsensors
to study the effects of salt concentration and temperature variation
on the conformation and motion of polythymine (polyT) strands of varying
lengths (10, 20, 50 nucleotides). PolyT strands were tethered to a
gold working electrode at the proximal end through a thiol linker
via covalent bonding between the Au electrode and sulfur link, which
can tend to decompose between a temperature range of 60 and 90 °C.
The strands were also modified with an electrochemically active methylene
blue (MB) moiety at the distal end. Electron transfer (eT) was measured
by square wave voltammetry (SWV) and used to infer information pertaining
to the average distance between the MB and the working electrode.
We observe changes in DNA flexibility due to varying ionic strength,
while the effects of increased DNA thermal motion are tracked for
elevated temperatures. This work elucidates the behavior of ssDNA
in the presence of a phosphate-buffered saline at NaCl concentrations
ranging from 20 to 1000 mmol/L through a temperature range of 10–50
°C in 1° increments, well below the decomposition temperature
range. The results lay the groundwork for studies on more complex
DNA strands in conjunction with different chemical and physical conditions.