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Applying a Linear Amplification Strategy to Recombinase Polymerase Amplification for Uniform DNA Library Amplification
- Source :
- ACS Omega, ACS Omega, Vol 4, Iss 22, Pp 19953-19958 (2019)
- Publication Year :
- 2019
- Publisher :
- American Chemical Society (ACS), 2019.
-
Abstract
- Recombinase polymerase amplification (RPA) is an isothermal DNA amplification method with broad applications as a point-of-care test and in molecular biology techniques. Currently, most of the applications are focused on target-specific amplification. Because RPA has the advantage of amplifying DNA under isothermal conditions, we utilized RPA as a DNA library amplification tool. In this study, we used a sheared genomic DNA library and an oligonucleotide (oligo) library for the comparison of polymerase chain reaction and RPA. For the sheared DNA library, we observed biased amplification after RPA was conducted. Thus, to amplify the size-variable DNA library uniformly, we introduced a linear amplification strategy with RPA and successfully improved the uniformity. On the other hand, using the same-sized oligo library, we confirmed that RPA amplified this library uniformly without modification of the protocol. These results demonstrate that RPA can be applied not only to amplify a specific target as previously demonstrated but also to amplify a complex DNA library composed of a large number of different DNA molecules.
- Subjects :
- Physics
Library
Oligonucleotide
General Chemical Engineering
Linear amplification
Recombinase Polymerase Amplification
General Chemistry
Computational biology
complex mixtures
Article
law.invention
Chemistry
genomic DNA
chemistry.chemical_compound
enzymes and coenzymes (carbohydrates)
chemistry
law
A-DNA
QD1-999
Polymerase chain reaction
DNA
Subjects
Details
- ISSN :
- 24701343
- Volume :
- 4
- Issue :
- 22
- Database :
- OpenAIRE
- Journal :
- ACS Omega
- Accession number :
- edsair.doi.dedup.....037ed0ceeb982c255435646e42007f2c
- Full Text :
- https://doi.org/10.1021/acsomega.9b02886