7 research outputs found
DNA Renaturation at the Water-Phenol Interface
We study DNA adsorption and renaturation in a water-phenol two-phase system,
with or without shaking. In very dilute solutions, single-stranded DNA is
adsorbed at the interface in a salt-dependent manner. At high salt
concentrations the adsorption is irreversible. The adsorption of the
single-stranded DNA is specific to phenol and relies on stacking and hydrogen
bonding. We establish the interfacial nature of a DNA renaturation at a high
salt concentration. In the absence of shaking, this reaction involves an
efficient surface diffusion of the single-stranded DNA chains. In the presence
of a vigorous shaking, the bimolecular rate of the reaction exceeds the
Smoluchowski limit for a three-dimensional diffusion-controlled reaction. DNA
renaturation in these conditions is known as the Phenol Emulsion Reassociation
Technique or PERT. Our results establish the interfacial nature of PERT. A
comparison of this interfacial reaction with other approaches shows that PERT
is the most efficient technique and reveals similarities between PERT and the
renaturation performed by single-stranded nucleic acid binding proteins. Our
results lead to a better understanding of the partitioning of nucleic acids in
two-phase systems, and should help design improved extraction procedures for
damaged nucleic acids. We present arguments in favor of a role of phenol and
water-phenol interface in prebiotic chemistry. The most efficient renaturation
reactions (in the presence of condensing agents or with PERT) occur in
heterogeneous systems. This reveals the limitations of homogeneous approaches
to the biochemistry of nucleic acids. We propose a heterogeneous approach to
overcome the limitations of the homogeneous viewpoint