Deposition of supercoiled DNA on mica for scanning force microscopy imaging.
The deposition of DNA molecules on mica is driven and controlled by the ionic densities around DNA and close to the surface of the substrate. Dramatic improvements in the efficiency and reproducibility of DNA depositions were due to the introduction of divalent cations in the deposition solutions. The ionic distributions on DNA and on mica determine the mobility of adsorbed DNA molecules, thus letting them assume thermodynamically equilibrated conformations, or alternatively trapping them in non-equilibrated conformations upon adsorption. With these prerequisites, mica does not seem like an inert substrate for DNA deposition for microscopy, and its properties greatly affect the efficiency of DNA deposition and the appearance of the molecules on the substrate. In our laboratory, we have some preliminary evidence that mica could also participate in DNA damage, most likely through its heavy metal impurities.
Samorì
B
B
Dipartimento di Chimica, Università della Calabria, Cosenza, Italy. samori@alma.unibo.it
Muzzalupo
I
I
Zuccheri
G
G
D000538:Aluminum Silicates; D002413:Cations, Divalent / Q000378:metabolism; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure*; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force / Q000379:methods*; D009690:Nucleic Acid Conformation; D013816:Thermodynamics
D000538:Aluminum Silicates; D002413:Cations, Divalent; D004278:DNA, Superhelical; C011934:mica