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Reduced Graphene Oxide-Based Solid-Phase Extraction for the Enrichment and Detection of microRNA
- Source :
- Analytical Chemistry. 89:10137-10140
- Publication Year :
- 2017
- Publisher :
- American Chemical Society (ACS), 2017.
-
Abstract
- MicroRNAs (miRNAs) are endogenous molecules with regulatory functions. The purification and enrichment of miRNA are essential for its precise and sensitive detection. miRNA isolated using commercial kits contains abundant interfering RNAs, and the concentration of miRNA may not be adequate for detection. Herein, we prepared a reduced graphene oxide (rGO)-based magnetic solid-phase extraction material for the enrichment and ultrasensitive detection of miRNA from intricate nucleic acid solutions. In situ reverse transcription (RT) was developed as the most efficient approach to desorb miRNA from rGO among the methods that are compatible for the subsequent amplification reported thus far. Additionally, rolling circle amplification and qPCR were used to detect let-7a with a decrease of the limit of detection by 24.7- and 31.3-fold, respectively. This material was also successfully used to extract and detect miRNA from total RNA isolated from human plasma. Our results show that the material prepared in this study has the potential for cancer biopsy in clinics and the discovery of new miRNAs in scientific research.
- Subjects :
- In situ
02 engineering and technology
Real-Time Polymerase Chain Reaction
01 natural sciences
Analytical Chemistry
Magnetics
Limit of Detection
microRNA
Humans
Solid phase extraction
DNA Primers
Detection limit
Chemistry
Solid Phase Extraction
010401 analytical chemistry
Nucleic acid amplification technique
021001 nanoscience & nanotechnology
Molecular biology
0104 chemical sciences
MicroRNAs
Real-time polymerase chain reaction
Biochemistry
Rolling circle replication
Nucleic acid
Graphite
0210 nano-technology
Nucleic Acid Amplification Techniques
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 89
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....80b82b48e7e4d8f4be005c4df818da5a