Open AccessCoupled Rolling Circle Amplification Loop-Mediated Amplification for Rapid Detection of Short DNA Sequences
Author(s) -
Jennifer Y. Marciniak,
Andrew C. Kummel,
Sadik C. Esener,
Michael J. Heller,
Bradley T. Messmer
Publication year - 2008
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112910
Subject(s) - rolling circle replication , loop mediated isothermal amplification , oligonucleotide , multiple displacement amplification , ligation , microbiology and biotechnology , dna , polymerase chain reaction , biology , base pair , recombinase polymerase amplification , d loop , multiplex ligation dependent probe amplification , dna polymerase , genetics , gene , dna extraction , mitochondrial dna , exon
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.
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