Bead-based padlock rolling circle amplification for single DNA molecule counting

Kae Sato; Reina Ishii; Naoki Sasaki; Kiichi Sato; Mats Nilsson

doi:10.1016/j.ab.2013.02.016

Bead-based padlock rolling circle amplification for single DNA molecule counting

Anal Biochem. 2013 Jun 1;437(1):43-5. doi: 10.1016/j.ab.2013.02.016. Epub 2013 Mar 1.

Authors

Kae Sato¹, Reina Ishii, Naoki Sasaki, Kiichi Sato, Mats Nilsson

Affiliation

¹ Department of Chemical and Biological Sciences, Japan Women's University, Bunkyou-ku, Tokyo, Japan. satouk@fc.jwu.ac.jp

PMID: 23467098
DOI: 10.1016/j.ab.2013.02.016

Abstract

Padlock rolling circle amplification (RCA) is a powerful analytical method for ultrasensitive DNA detection. Although there are some advantages to bead-based RCA, a detailed study of the relationship between the bead material and the efficiency of bead-based RCA has not been reported. Here, we compared the reaction efficiencies of bead-based RCA performed on two types of bead material: agarose and polystyrene. Agarose was a more suitable material for on-bead RCA. The calibration curve showed linearity between 0.05 and 1 nM, and the limit of detection was 9 pM (9 amol) for Salmonella DNA determination.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
DNA, Bacterial / analysis*
DNA, Bacterial / genetics
Microspheres*
Nucleic Acid Amplification Techniques / methods*
Nucleic Acid Hybridization
Salmonella / genetics

Substances

DNA, Bacterial