Attomole DNA detection assay via rolling circle amplification and single molecule detection.
A bead-based assay was developed for highly sensitive single molecule DNA detection. Rolling circle amplification (RCA), an isothermal amplification technique that creates tandem repeated sequences, was used in combination with a fluorescent complementary DNA to create dense clusters of fluorescence. These clusters, each corresponding to a single target molecule, can be detected unambiguously due to their high signal/noise ratios. The limit of detection of this assay is approximately 1 amol. This simple single molecule assay allows high detection sensitivity without the use of complex equipment.
Schopf
Eric
E
Department of Mechanical and Aerospace Engineering, California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. eschopf@ucla.edu
Chen
Yong
Y
D004247:DNA / Q000032:analysis* / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012685:Sepharose / Q000737:chemistry
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA; D012685:Sepharose