Molecular crowding improves bead-based padlock rolling circle amplification

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• 作者：Naoki Sasaki^a ; ^{nsasaki@toyo.jp} ; Yoshitaka Gunji^a ; Chikako Kase^b ; Kae Sato^b
• 关键词：Padlock probe ; Rolling circle amplification ; Microbeads ; Molecular crowding ; Poly (ethylene glycol) ; Single nucleotide polymorphism
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：15 February 2017
• 年：2017
• 卷：519
• 期：Complete
• 页码：15-18
• 全文大小：796 K
• 卷排序：519

文摘

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.