Overview

Profile across the rat miRNome

SBI has simplified qPCR profiling of rat miRNAs by bringing together a comprehensive qPCR array of rat miRNAs. Our Rat rno-miRNome MicroRNA Profiling Kit comes with all the reagents necessary to tag and convert small RNAs into quantifiable cDNA using the sensitive QuantiMir™ technology and a large set of rat miRNA qPCR assays, including major and minor miR forms (723 mature miRNAs).

The kits include assays in two, pre-formatted 384-well assay plates with three endogenous reference miRNA controls on each plate (U1, U6, and RNU43). All microRNA assays are based on miRBase entries.

• Identify miRNA biomarkers and expression pattern signatures
• Rapidly tag and convert all small RNAs into detectable cDNA for qPCR
• Measure as little as picogram amounts of starting total RNA
• Conduct high-throughput screens of cell lines, tissues, and clinical samples

About the included QuantiMir Kit

When you’d like to profile miRNAs with qPCR, SBI’s QuantiMir™ Kit can get your samples ready. By efficiently converting all of your miRNA into cDNA, you can get accurate, sensitive, and quantitative qPCR data on the miRs in your sample. Great for understanding differential miRNA expression, simultaneously profiling siRNA and mRNA, and more.

• Simple and robust procedure
• Rapidly and efficiently convert all small RNAs into cDNAs for qPCR
• Suitable for high-throughput screening of clinical samples
• Sensitive and accurate
• Versatile—design your own miRNA assays

How It Works

Using the rno-miRNome MicroRNA Profiling Kit

First, prepare RNA for qPCR using the QuantiMir Kit

Highly efficient poly-A tailing and reverse transcription in a single reaction tube provides uniform cDNA synthesis of miRNAs. The optimized reaction conditions and buffer components maximize cDNA yield when starting with several micrograms down to picograms of input total RNA. The universal 3′ tag sequence incorporated during reverse transcription enables easily scalable and accurate miRNA expression analysis by qPCR—profile thousands of different miRNAs from a single reverse transcription reaction.

• Tag all small RNAs with a poly-A tail
• Anneal an oligo-dT adaptor
• Reverse transcribe to create first-strand cDNA

The result is a cDNA pool of anchor-tailed miRNAs that are ready for qPCR.

Then conduct qPCR assays

Get additional information about the array including miRNA sequences, miRbase accession numbers, and qPCR analysis templates in the Excel document, “Free Analysis Software for Rat miRNome Ver. 19”

Supporting Data

miRNome Profiler Performance Data

Figure 1. Accurately normalize your microRNA expression profiling data using three reference controls—U6 snRNA, RNU1A, and RNU43.

Figure 2. Get high reproducibility—excellent technical replicates across 8-logs. Reliable replicates from plate-to-plate enable precise comparisons of numerous samples across large microRNA profiling studies.

Resources

Citations

• Olivieri, F, et al. (2016) Circulating miRNAs and miRNA shuttles as biomarkers: Perspective trajectories of healthy and unhealthy aging. Mech. Ageing Dev..2016 Dec 13;. PM ID:27986629
• Khamaisi, M, et al. (2016) PKCδ inhibition normalizes the wound-healing capacity of diabetic human fibroblasts. J. Clin. Invest..2016 Mar 1; 126(3):837-53. PM ID:26808499
• Pandey, S, et al. (2015) The RNA Stem-Loop to G-Quadruplex Equilibrium Controls Mature MicroRNA Production inside the Cell. Biochemistry.2015 Dec 8; 54(48):7067-78. PM ID:26554903
• Meiler, S, et al. (2015) MicroRNA 302a is a novel modulator of cholesterol homeostasis and atherosclerosis. Arterioscler. Thromb. Vasc. Biol..2015 Feb 1; 35(2):323-31. PM ID:25524771
• 岡本行広, , et al. (2015) マイクロ抽出場によるヒト腫瘍細胞由来のマイクロ RNA 抽出法. 分析化学.2015 Jan 31; 64(1):9-13. Link:分析化学
• Kumarswamy, R, et al. (2012) SERCA2a gene therapy restores microRNA-1 expression in heart failure via an Akt/FoxO3A-dependent pathway. Eur. Heart J..2012 May 1; 33(9):1067-75. PM ID:22362515
• Alpini, G, et al. (2011) Regulation of placenta growth factor by microRNA-125b in hepatocellular cancer. J. Hepatol..2011 Dec 1; 55(6):1339-45. PM ID:21703189
• Crother, TR, et al. (2011) Chlamydia pneumoniae infection induced allergic airway sensitization is controlled by regulatory T-cells and plasmacytoid dendritic cells. PLoS ONE.2011 Jun 22; 6(6):e20784. PM ID:21695198
• Stellato, C, et al. (2011) Glucocorticoid (GC) Modulation of Global miRNA Profile in Human Airway Epithelial Cells. Journal of Allergy and Clinical Immunology.2011 Feb 1; 127(2S):AB64. Link:Journal of Allergy and Clinical Immunology
• Karim, Z, et al. (2011) Signaling Mechanism of Tolerance following Repetitive Stimulation with a Single Allergen and its Reversal by Stimulation with Multiple Allergens. Journal of Allergy and Clinical Immunology.;:AB64. Link:Journal of Allergy and Clinical Immunology