| Chapter title |
Detection and Quantification of MicroRNAs by Ligase-Assisted Sandwich Hybridization on a Microarray.
|
|---|---|
| Chapter number | 5 |
| Book title |
Microarray Technology
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3136-1_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3135-4, 978-1-4939-3136-1
|
| Authors |
Iizuka, Ryo, Ueno, Taro, Funatsu, Takashi, Ryo Iizuka, Taro Ueno, Takashi Funatsu |
| Abstract |
Extracellular microRNAs (miRNAs) in body fluids have been identified as promising biomarkers for different human diseases. The high-throughput, multiplexed detection and quantification of these miRNAs are highly beneficial for the rapid and accurate diagnosis of diseases. Here, we developed a simple and convenient microarray-based technique, named ligase-assisted sandwich hybridization (LASH), for the detection and quantification of miRNAs. The LASH assay involves the hybridization of capture and detection probe pairs with the target miRNA to form a double-stranded structure which is then nick-sealed by T4 DNA ligase. Using this assay, we successfully demonstrated the multiplexed detection and quantification of different miRNAs in total RNA samples derived from blood obtained within 3 h. Here, we provide a detailed protocol for the LASH assay to detect a specific miRNA, as a model for the detection and quantification of extracellular miRNAs. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 2 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 50% |
| Members of the public | 1 | 50% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 7 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 2 | 29% |
| Unspecified | 1 | 14% |
| Professor | 1 | 14% |
| Other | 1 | 14% |
| Student > Master | 1 | 14% |
| Other | 1 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 43% |
| Agricultural and Biological Sciences | 2 | 29% |
| Unspecified | 1 | 14% |
| Engineering | 1 | 14% |