Direct atomic force microscopy observations of monovalent ion induced binding of DNA to mica.
Multivalent ions in solution are known to mediate attraction between two like-charged molecules. Such attraction has proved useful in atomic force microscopy (AFM) where DNA may be immobilized to a mica surface facilitating direct imaging in liquid. Theories of DNA immobilization suggest that either 'salt bridging' or fluctuation in the positions of counter ions about both the mica surface and DNA backbone secure DNA to the mica substrate. Whilst both theoretical and experimental evidence suggest that immobilization is possible in the presence of divalent ions, very few studies identify that such immobilization is possible with monovalent ions. Here we present direct AFM evidence of DNA immobilized to mica in the presence of only monovalent ions. Our data depict E. coli plasmid pBR322 adsorbed onto the negatively charged mica both after short (10 min) and long (24 h) incubation periods. These data suggest the need to re-explore current theories of like-charge attraction to include the possibility of monovalent interactions. We suggest that this DNA immobilization strategy may offer the potential to image natural processes with limited immobilization forces and hence enable maximum conformational freedom of the immobilized biomolecule.
Laboratory of Biophysics and Surface Analysis, School of Pharmacy, The University of Nottingham, Nottingham NG7 2RD, UK.
J S
JS
Ellis
H G
HG
Abdelhady
S
S
Allen
M C
MC
Davies
C J
CJ
Roberts
S J B
SJ
Tendler
P M
PM
Williams
D000538:Aluminum Silicates / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D018625:Microscopy, Atomic Force / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D012964:Sodium / Q000494:pharmacology*
D000538:Aluminum Silicates; D004269:DNA, Bacterial; D012964:Sodium; C011934:mica