Immobilization and hybridization of oligonucleotides on maleimido-terminated self-assembled monolayers.
Maleimido-terminated self-assembled monolayers were prepared via a one-step reaction of a maleimido-functionalized trichlorosilane with an oxide-covered surface. Through conjugate addition, thiol-tagged DNA was immobilized on these maleimido-functionalized self-assembled monolayers, and these immobilized oligonucleotides were further hybridized with complementary strands. The surface concentrations of oligonucleotides were quantified with radioactivity measurements. The surface concentrations of immobilized oligonucleotides increase in a nonlinear manner with increasing molar fraction of maleimido moieties on the surface and achieve high values of 1.6 (+/-0.6) x 10(13) oligonucleotides/cm2 if monolayers having full coverage of maleimido groups are used. High hybridization efficiencies of complementary strands (approximately 75%) are obtained even if the surface concentration of immobilized strands exceeds a value of 5.0 x 10(12) oligonucleotides/cm2. Furthermore, five cycles of denaturation and rehybridization result in only a 4% loss of the immobilized DNA after each run.
Wang
Yayun
Y
Department of Organic and Macromolecular Chemistry, OCIII, University of Ulm, Ulm 89069, Germany.
Prokein
Tobias
T
Hinz
Michael
M
Seliger
Hartmut
H
Goedel
Werner A
WA
D008301:Maleimides / Q000737:chemistry*; D008567:Membranes, Artificial; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry*; D012821:Silanes / Q000737:chemistry*; D013055:Spectrophotometry, Infrared
D008301:Maleimides; D008567:Membranes, Artificial; D009841:Oligonucleotides; D012821:Silanes; C506921:maleimido-undecyltrichlorosilane; C065893:trichlorosilane