Coupled rolling circle amplification loop-mediated amplification for rapid detection of short DNA sequences.
Circularizable oligonucleotide probes can detect short DNA sequences with single-base resolution at the site of ligation and can be amplified by rolling circle amplification (RCA) using strand displacing polymerases. A secondary amplification scheme was developed that uses the loop-mediated amplification reaction concurrent with RCA to achieve rapid signal development from the starting circular molecules. This isothermal reaction was found to be significantly faster than the comparable hyperbranching amplification method and could detect 100 circular copies in less than 1 h.
Marciniak
Jennifer
J
Department of Bioengineering, University of California-San Diego, La Jolla, CA, USA.
Kummel
Andrew
A
Esener
Sadik
S
Heller
Michael
M
Messmer
Bradley
B
D001483:Base Sequence; D011088:DNA Ligases / Q000378:metabolism; D015342:DNA Probes / Q000235:genetics; D004270:DNA, Circular / Q000737:chemistry / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics*; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
D015342:DNA Probes; D004270:DNA, Circular; D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes; D011088:DNA Ligases