The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.
Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs
Open accessAbstract
MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 mul of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.
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Bartel (2004) · doi:10.1016/s0092-8674(04)00045-5 · Cited by 34,714
Cleavage of Scarecrow-like mRNA Targets Directed by a Class of Arabidopsis miRNA
Llave, C., Xie, Z., Kasschau, K. D., & Carrington, J. C. (2002) · Science · doi:10.1126/science.1076311 · Cited by 1,627
Specific effects of microRNAs on the plant transcriptome
Schwab, R., Palatnik, J. F., Riester, M., Schommer, C., Schmid, M., & Weigel, D. (2005) · Developmental Cell · doi:10.1016/j.devcel.2005.01.018 · Cited by 1,467
Cited by
Benes, V., & Castoldi, M. (2010) · Methods · doi:10.1016/j.ymeth.2010.01.026 · Cited by 361
Let-7 microRNA–mediated regulation of IL-13 and allergic airway inflammation
Kumar, M., Ahmad, T., Sharma, A., Mabalirajan, U., Kulshreshtha, A., Agrawal, A., & Ghosh, B. (2011) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2011.04.034 · Cited by 310
Ryoo, S.-R., Lee, J., Yeo, J., Na, H.-K., Kim, Y.-K., Jang, H., Lee, J. H., Han, S. W., Lee, Y., Kim, V. N., et al. (2013) · ACS Nano · doi:10.1021/nn401183s · Cited by 309
Li, H., Zhang, Z., Huang, F., Chang, L., & Ma, Y. (2009) · Plant Cell Reports · doi:10.1007/s00299-009-0693-3 · Cited by 46
Tissue Culture Responsive MicroRNAs in Strawberry
Li, H., Zhao, X., Dai, H., Wu, W., Mao, W., & Zhang, Z. (2012) · Plant Molecular Biology Reporter · doi:10.1007/s11105-011-0406-2 · Cited by 29
Metadata
- Authors
- Varkonyi-Gasic, E., Wu, R., Wood, M., Walton, E. F., & Hellens, R. P.
- Venue
- Plant Methods
- Published
- 2007
- Volume
- 3
- Issue
- 1
- Pages
- 12
- ISSN
- 1746-4811 (ISSN-L)
- OpenAlex
- W1966612206
- Cited by
- 1,186
- References
- 3
- FWCI
- 20.79
- Type
- journal article
- Language
- en
- Open access
- gold
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- Varkonyi-Gasic, E., Wu, R., Wood, M., Walton, E. F., & Hellens, R. P. (2007). Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant Methods. https://doi.org/10.1186/1746-4811-3-12
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