The DOI matches its Crossref record; 9 of the 9 references it cites were found in a registry or an index.
Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification
Open accessAbstract
MicroRNAs are a class of small non-coding RNAs that serve as important regulators of gene expression at the posttranscriptional level. They are stable in body fluids and pose great potential to serve as biomarkers. Here, we present a highly specific, sensitive and cost-effective system to quantify miRNA expression based on two-step RT-qPCR with SYBR-green detection chemistry called Two-tailed RT-qPCR. It takes advantage of novel, target-specific primers for reverse transcription composed of two hemiprobes complementary to two different parts of the targeted miRNA, connected by a hairpin structure. The introduction of a second probe ensures high sensitivity and enables discrimination of highly homologous miRNAs irrespectively of the position of the mismatched nucleotide. Two-tailed RT-qPCR has a dynamic range of seven logs and a sensitivity sufficient to detect down to ten target miRNA molecules. It is capable to capture the full isomiR repertoire, leading to accurate representation of the complete miRNA content in a sample. The reverse transcription step can be multiplexed and the miRNA profiles measured with Two-tailed RT-qPCR show excellent correlation with the industry standard TaqMan miRNA assays (r2 = 0.985). Moreover, Two-tailed RT-qPCR allows for rapid testing with a total analysis time of less than 2.5 hours.
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Cites
MicroRNAs: Target Recognition and Regulatory Functions
Bartel, D. P. (2009) · Cell · doi:10.1016/j.cell.2009.01.002 · Cited by 20,068
The widespread regulation of microRNA biogenesis, function and decay
Krol, J., Loedige, I., & Filipowicz, W. (2010) · Nature Reviews Genetics · doi:10.1038/nrg2843 · Cited by 4,672
Gene silencing by microRNAs: contributions of translational repression and mRNA decay
Huntzinger, E., & Izaurralde, E. (2011) · Nature Reviews Genetics · doi:10.1038/nrg2936 · Cited by 2,339
LNA (Locked Nucleic Acid): High-Affinity Targeting of Complementary RNA and DNA
Vester, B., & Wengel, J. (2004) · Biochemistry · doi:10.1021/bi0485732 · Cited by 727
IsomiRs – the overlooked repertoire in the dynamic microRNAome
Neilsen, C. T., Goodall, G. J., & Bracken, C. P. (2012) · Trends in Genetics · doi:10.1016/j.tig.2012.07.005 · Cited by 499
Properties of the Reverse Transcription Reaction in mRNA Quantification
Ståhlberg, A., Håkansson, J., Xian, X., Semb, H., & Kubista, M. (2004) · Clinical Chemistry · doi:10.1373/clinchem.2003.026161 · Cited by 409
Self-citationMicroRNAs and their isomiRs function cooperatively to target common biological pathways
Cloonan, N., Wani, S., Xu, Q., Gu, J., Lea, K., Heater, S., Barbacioru, C., Steptoe, A. L., Martin, H. C., Nourbakhsh, E., et al. (2011) · Genome Biology · doi:10.1186/gb-2011-12-12-r126 · Cited by 345
Comparison of Reverse Transcriptases in Gene Expression Analysis
Ståhlberg, A., Kubista, M., & Pfaffl, M. (2004) · Clinical Chemistry · doi:10.1373/clinchem.2004.035469 · Cited by 240
Self-citationSingle-cell sequencing of the small-RNA transcriptome
Faridani, O. R., Abdullayev, I., Hagemann-Jensen, M., Schell, J. P., Lanner, F., & Sandberg, R. (2016) · Nature Biotechnology · doi:10.1038/nbt.3701 · Cited by 212
Cited by
Tian, T., Shu, B., Jiang, Y., Ye, M., Liu, L., Guo, Z., Han, Z., Wang, Z., & Zhou, X. (2021) · ACS Nano · doi:10.1021/acsnano.0c08165 · Cited by 328
Dave, V. P., Ngo, T. A., Pernestig, A.-K., Tilevik, D., Kant, K., Nguyen, T., Wolff, A., & Bang, D. D. (2019) · Laboratory Investigation · doi:10.1038/s41374-018-0143-3 · Cited by 248
Wright, C., Rajpurohit, A., Burke, E. E., Williams, C., Collado-Torres, L., Kimos, M., Brandon, N. J., Cross, A. J., Jaffe, A. E., Weinberger, D. R., et al. (2019) · BMC Genomics · doi:10.1186/s12864-019-5870-3 · Cited by 99
Metadata
- Authors
- Androvic, P., Valihrach, L., Elling, J., Sjoback, R., & Kubista, M.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2017-07-13
- Volume
- 45
- Issue
- 15
- Pages
- e144-e144
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2734814984
- Cited by
- 198
- References
- 9
- FWCI
- 5.53
- Type
- journal article
- Language
- en
- Open access
- gold
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- Androvic, P., Valihrach, L., Elling, J., Sjoback, R., & Kubista, M. (2017). Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification. Nucleic Acids Research. https://doi.org/10.1093/nar/gkx588
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