Chapter title |
Quantitative Real-Time PCR Analysis of MicroRNAs and Their Precursors Regulated by TGF-β Signaling.
|
---|---|
Chapter number | 20 |
Book title |
TGF-β Signaling
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-2966-5_20 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2965-8, 978-1-4939-2966-5
|
Authors |
Kang, Hara, Hata, Akiko, Hara Kang, Akiko Hata |
Abstract |
The signaling pathway of TGF-β and its family member BMP has been implicated in vascular development and maintenance of homeostasis by modulating expression of small noncoding microRNAs (miRNAs). MiRNAs repress target genes, which play a critical role in regulating vascular smooth muscle cell (VSMC) growth, phenotype, and function. To understand the mechanisms by which specific miRNAs control the TGF-β and BMP signaling pathway in VSMC, it is essential to quantitate levels of specific miRNAs and their precursors whose expression are controlled by TGF-β/BMP signaling. Here, we describe a real-time quantization method for accurate and sensitive detection of miRNAs and their precursors, such as primary transcripts of miRNAs (pri-miRNAs) and precursor miRNAs (pre-miRNAs). This method requires two steps; synthesis of single-stranded complementary DNAs (cDNAs) from total RNA samples and quantization of specific pri-, pre-, or mature miRNAs by quantitative polymerase chain reaction (PCR) using a real-time PCR machine. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 6 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 3 | 50% |
Professor | 1 | 17% |
Student > Bachelor | 1 | 17% |
Student > Ph. D. Student | 1 | 17% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 33% |
Agricultural and Biological Sciences | 2 | 33% |
Environmental Science | 1 | 17% |
Immunology and Microbiology | 1 | 17% |