Two-Tailed Reverse Transcription
Frozen. Upon receipt, store the product at the temperature recommended below.
Store the kit at -20°C. Under these conditions, assay components are stable till the expiry date is over. (See the expiry date indicated on the kit label).
Below, links to independent papers developing and using Two-Tailed PCR can be found
- Androvic P, Valihrach L, Elling J, Sjoback R, Kubista M. Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification. Nucleic Acids Res. 2017 Sep 6;45(15):e144. doi: 10.1093/nar/gkx588. PMID: 28911110; PMCID: PMC5587787. See more on PubMed
- Damayanti F, Lombardo F, Masuda JI, Shinozaki Y, Ichino T, Hoshikawa K, Okabe Y, Wang N, Fukuda N, Ariizumi T, Ezura H. Functional Disruption of the Tomato Putative Ortholog of HAWAIIAN SKIRT Results in Facultative Parthenocarpy, Reduced Fertility and Leaf Morphological Defects. Front Plant Sci. 2019 Oct 14;10:1234. doi: 10.3389/fpls.2019.01234. PMID: 31681360; PMCID: PMC6801985. See more on PubMed
- Anna BB, Grzegorz B, Marek K, Piotr G, Marcin F. Exposure to High-Intensity Light Systemically Induces Micro-Transcriptomic Changes in Arabidopsis thaliana Roots. Int J Mol Sci. 2019 Oct 16;20(20):5131. doi: 10.3390/ijms20205131. PMID: 31623174; PMCID: PMC6829545. See more on PubMed
- Click here for more papers citing Two-tailed PCR
- For research use only.
- The total assay time is less than 1 hours.
- The kit measures specific miRNA in biological fluids.
- Components of the kit are provided ready to use.
Exogenous control, Spike-in control
MicroRNAs (miRNAs) are small non-coding RNA molecules, approximately 22 nucleotides in length that regulate gene translation through silencing or degradation of target mRNAs. They are involved in multiple biological processes, including differentiation and proliferation, metabolism, hemostasis, apoptosis or inflammation, and in the pathophysiology of many diseases. Numerous studies have suggested circulating miRNAs as promising diagnostic and prognostic biomarkers of many diseases.
Concentration of target miRNA measured by miREIA can be affected by efficiency of RNA isolation. Therefore, it is recommended to normalize the measured concentration of target miRNA to the efficiency of RNA isolation using an exogenous control.
Efficiency of RNA isolation can be monitored by adding a defined amount of synthetic non-human miRNA, e.g. cel-miR-39-3p miRNA, to the sample during RNA isolation. The synthetic RNA is processed the same way as the target RNA present in the samples.
After RNA isolation, concentration of exogenous cel-miR-39-3p added to the samples is measured by cel-miR-39-3p miREIA in parallel with the concentration of the target miRNA. To calculate the coefficient of isolation efficiency, the amount of cel-miR-39-3p added to the samples prior to isolation is divided by the concentration of cel-miR-39-3p measured by miREIA.
Finally, the concentration of target miRNA measured by miREIA is multiplied by the coefficient of isolation efficiency for every sample.
If such a normalization is applied, then, each RNA isolate sample is measured by two miREIA kits: one kit for determination of the target miRNA (samples are diluted as recommended in the corresponding datasheet) and by a second kit, cel-miR-39-3p miREIA (samples are diluted 10-fold as written in page 15).