Chapter title |
Stem-Loop qRT-PCR for the Detection of Plant microRNAs.
|
---|---|
Chapter number | 13 |
Book title |
Plant Epigenetics
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4899-7708-3_13 |
Pubmed ID | |
Book ISBNs |
978-1-4899-7706-9, 978-1-4899-7708-3
|
Authors |
Erika Varkonyi-Gasic |
Editors |
Igor Kovalchuk |
Abstract |
Plant microRNAs (miRNAs) play important roles in the posttranscriptional regulation of protein-coding genes, and they are essential for a normal development and survival. Mature miRNAs are cleaved from larger precursor RNAs and are typically 21-22 nt long.The small size, the lack of a common feature like a poly(A) tail, 3' end-modifications, and presence of a precursor-all these factors affect the detection and hinder the quantification of miRNAs. The stem-loop qRT-PCR method described here is designed to detect and quantify mature miRNAs in a fast, specific, accurate, and reliable manner. Firstly, a miRNA-specific stem-loop RT primer is hybridized to miRNA and then reverse transcribed. Next, the RT product is amplified and monitored in real time using a miRNA-specific forward primer and a universal reverse primer. This method enables miRNA expression profiling from as little as 10 pg of total RNA, and it is suitable for a relatively high-throughput analysis of miRNA expression. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 38 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 9 | 24% |
Researcher | 7 | 18% |
Student > Master | 5 | 13% |
Student > Bachelor | 4 | 11% |
Other | 3 | 8% |
Other | 1 | 3% |
Unknown | 9 | 24% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 14 | 37% |
Biochemistry, Genetics and Molecular Biology | 9 | 24% |
Medicine and Dentistry | 1 | 3% |
Neuroscience | 1 | 3% |
Engineering | 1 | 3% |
Other | 0 | 0% |
Unknown | 12 | 32% |