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Visual Detection of Multiple Nucleic Acids in a Capillary Array
Published on: November 15, 2017
Coupled rolling circle amplification loop-mediated amplification for rapid detection of short DNA sequences
Jennifer Marciniak1, Andrew Kummel, Sadik Esener
1Department of Bioengineering, University of California-San Diego, La Jolla, CA, USA.
Biotechniques
|September 10, 2008
Summary
Circularizable oligonucleotide probes detect DNA with single-base precision. A novel amplification method combining rolling circle amplification and loop-mediated amplification offers rapid, sensitive DNA detection.
Area of Science:
- Molecular Biology
- Biotechnology
- Genetics
Background:
- Circularizable oligonucleotide probes offer precise detection of short DNA sequences.
- Rolling circle amplification (RCA) is a method for amplifying circular DNA molecules.
Purpose of the Study:
- To develop a rapid and sensitive DNA detection method.
- To enhance signal development from circular oligonucleotide probes.
Main Methods:
- Utilized circularizable oligonucleotide probes for DNA detection.
- Developed a secondary amplification scheme combining loop-mediated amplification (LAMP) with RCA.
- Employed strand-displacing polymerases for isothermal amplification.
Main Results:
- Achieved rapid signal development from circular molecules.
- The combined RCA and LAMP method was significantly faster than hyperbranching amplification.
- Detected 100 circular copies in under 1 hour.
Conclusions:
- The developed isothermal amplification scheme provides a fast and sensitive method for DNA detection.
- This technique enhances the utility of circularizable oligonucleotide probes.
- Offers a potential advancement in molecular diagnostics and research.
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