title: Molecular crowding improves bead-based padlock rolling circle amplification.
abstract: Bead-based padlock rolling circle amplification (RCA), an ultrasensitive and accurate DNA detection technique, was conducted in a molecular crowding environment created by poly(ethylene glycol) (PEG). The number of RCA products generated increased and exhibited a bell-shaped dependence on PEG concentration. Experiments using magnetic beads suggested that facilitation of DNA ligation and hybridization is the main reason for the observed increase. Selectivity of the technique was retained in the presence of PEG. This technique is simple and can be utilized to detect target DNA with high accuracy and sensitivity in a variety of areas such as medical diagnosis and food analysis.
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Toyo University, 2100 Kujirai, Kawagoe, Saitama 350-8585, Japan. Electronic address: nsasaki@toyo.jp.
authors.forename: Naoki
authors.initials: N
authors.lastname: Sasaki
authors.affiliation: Department of Applied Chemistry, Faculty of Science and Engineering, Toyo University, 2100 Kujirai, Kawagoe, Saitama 350-8585, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Gunji
authors.affiliation: Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Kase
authors.affiliation: Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan.
authors.forename: Kae
authors.initials: K
authors.lastname: Sato
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*
chemical_list: D011092:Polyethylene Glycols; D004247:DNA
keywords: Microbeads; Molecular crowding; Padlock probe; Poly (ethylene glycol); Rolling circle amplification; Single nucleotide polymorphism