title: Enzymatic preparation of multimilligram amounts of pure single-stranded DNA samples for material and analytical sciences.
abstract: We present a method for high-yield production of multimilligram amounts of pure single-stranded DNA employing rolling circle amplification (RCA) and processing by restriction enzymes. Pure and homogeneous samples are produced with minimal handling time, reagents, and waste products. The RCA method is more than twice as efficient in dNTP incorporation than conventional polymerase chain reaction in producing end product. The validity and utility of the method are demonstrated in the production of a uniformly (13)C/(15)N-labeled 38-nt cocaine aptamer DNA used in nanosensing devices.
authors: Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
authors: Frank H T
authors: FH
authors: Nelissen
authors: Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
authors: Elles P M
authors: EP
authors: Goossens
authors: Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
authors: Marco
authors: M
authors: Tessari
authors: Department of Biophysical Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands. Electronic address: h.heus@science.ru.nl.
authors: Hans A
authors: HA
authors: Heus
mesh_terms: D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D003042:Cocaine / Q000032:analysis; D004277:DNA, Single-Stranded / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D003042:Cocaine
keywords: DNA aptamer; DNA synthesis; Isotope labeling; NMR; Rolling circle amplification