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Microrna specific primers

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MicroRNA-specific primers are oligonucleotide sequences designed to selectively amplify and detect specific microRNA molecules. They are used in molecular biology and genomics research to study the expression and function of microRNAs.

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Lab products found in correlation

Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Rneasy kit, by Qiagen (1 mentions) Abi prism 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions) Rq manager software v1, by Thermo Fisher Scientific (1 mentions) Taqman microrna assay kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

MicroRNAs species-specific primers TaqMan microRNA-specific primers MicroRNA-specific stem-loop primers Specific primers

3 protocols using microrna specific primers

1

Cardiac RNA Extraction and cDNA Synthesis

Cited 2 times
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RNA was extracted using Qiagen RNeasy kits (Qiagen, Hilden, Germany) from the viable myocardium of the remote noninfarct region of the left ventricle (30 mg) from MI rats. This region of the myocardium was chosen because it receives even exposure to uremic toxins concentrations observed in the circulation. A similar region of the heart was obtained for sham rats (Lekawanvijit et al. 2013 (link)). Then cDNA synthesis was performed using microRNA-specific primers (Applied Biosystems, Foster City, CA), by method provided by the manufacturer. Standard cDNA synthesis protocol was followed to obtain cDNA for gene assays (Nankervis et al. 2013 (link)).
Rana I., Kompa A.R., Skommer J., Wang B.H., Lekawanvijit S., Kelly D.J., Krum H, & Charchar F.J. (2015). Contribution of microRNA to pathological fibrosis in cardio-renal syndrome: impact of uremic toxins. Physiological Reports, 3(4), e12371.
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2

Quantitative microRNA Expression Analysis

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MicroRNA expression was determined by real time reverse transcriptase polymerase chain reaction (real time RT-PCR). Complementary DNA (cDNA) was made from 10ng of total RNA from each sample using the TaqMan micro RNA reverse transcription kit (Applied Biosystems, Foster City, CA) and microRNA specific primers for miR-29a, miR-29b, and miR-29c (Applied Biosystems). These cDNA samples, along with RNU48 as a loading control, were then amplified using the ABI TaqMan MiRNA PCR Kit (Applied Biosystems) and the ABI Prism 7900HT sequence detection system (Applied Biosystems). The real time PCR was performed in triplicate in a minimum of three subjects per experiment. Values for each microRNA were normalized to the expression levels of RNU 48 using 2−ΔΔCT methodology (22 (link)).
MARSH E.E., STEINBERG M.L., PARKER J.B., WU J., CHAKRAVARTI D, & BULUN S.E. (2016). Decreased expression of MicroRNA-29 family in leiomyoma contributes to increased major fibrillar collagen production. Fertility and sterility, 106(3), 766-772.
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3

Validation of Microarray miRNA Expression

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Microarray expression of selected microRNAs was validated by RT-qPCR, both in the same cohort and in the validation cohort as described. The RT-qPCR was done using a microRNA-specific TaqMan microRNA assay kit (Applied Biosystems, Foster City, CA) and Applied Biosystems 7900H Fast Real-Time system. Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table S2).
The cDNA from the reverse transcription reaction was added to the TaqMan Universal Master Mix II no UNG and the labeled microRNA specific TaqMan probes (Applied Biosystems). RNU 6B, small nuclear RNAs, were used as endogenous controls for normalization of input cDNA levels (Applied Biosystems). We used a commercial Ambion breast control (Life Technologies, http://www.lifetechnologies.com/) as a calibrator.
The RT-qPCR was performed on all samples in duplicate. MicroRNA expression was quantified as ΔΔCt values, where Ct = threshold cycle, ΔCt = (Ct target microRNA − Ct RNU 6B) and ΔΔCt = (ΔCt target microRNA − ΔCt calibrator), and fold change for each microRNA was calculated by the method.17 (link) This was done using RQ manager software, v. 1.2 (Applied Biosystems).
Bjaanæs M.M., Halvorsen A.R., Solberg S., Jørgensen L., Dragani T.A., Galvan A., Colombo F., Anderlini M., Pastorino U., Kure E., Børresen-Dale A.L., Brustugun O.T, & Helland Å. (2014). Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas. International Journal of Cancer. Journal International du Cancer, 135(8), 1812-1821.
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