Overview
Profile regulatory RNAs for deeper insights into cellular processes
Profile 89 regulatory RNAs with this kit that includes reagents for converting RNAs to cDNAs and a primer set for qPCR assays in a 96-well format. All primers have been validated against total RNA from several cell lines and tissues such as HEK293, HT1080, MCF7 (human breast cancer cell line), human fibroblast cells, human fetal brain, human adult brain, iPSC cells, and neural precursor cells (NPCs).
qPCR assays marked in blue are for eRNAs, green for lncRNAs and controls are indicated in black.
The kit includes four RNA controls for normalization—GAPDH, Actin, U6, and 18SrRNA—as well as assays for FOXO3 and BRD4 to detect eRNAs.
How It Works
Using the Regulatory RNA qPCR Profiler
First, get your regulatory RNA ready for qPCR by converting to cDNA
While some lncRNAs have endogenouse polyA tails, other lncRNAs do not. To enhance qPCR assay performance, the cDNA synthesis workflow includes steps and reagents to polyadenylate all lncRNAs before cDNA conversion using the tagged oligo dT adaptor and random primers. This combined RNA tailing and oligo dT plus random primers boosts cDNA yield significantly and enables strand-specific lncRNA qPCR profiling.
- Tag all lncRNAs 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 lncRNAs that are ready for qPCR.
Then conduct qPCR assays
Get additional information about the array including RNA sequences, database links, and qPCR analysis templates in the Excel document, “Assay List and Free Analysis Software for Human Regulatory RNA qPCR Array ”
Supporting Data
Example Regulatory RNA qPCR Profiler Data
Figure 1. Sample dissociation analysis curves.
Figure 2. Profile regulatory RNAs in cancer. Representative eRNAs and LncRNAs that are significantly up regulated in cancer cells. RNA from HFF (normal) and HT1080 (cancer) cells were purified, converted to cDNA and the cDNA was profiled for expression using the regulatory RNA profiler.
Figure 3. Detect regulatory RNAs in exosomes. After isolating exosomes using ExoQuick (Cat.# EXOQ5A-1) and then exosomal RNAs using SeraMir (Cat.# RA800A-1), we converted the exosomal RNA to cDNA using the Exosomal RNA was converted to cDNA using the RNA-Quant kit (Cat.# RA430A-1) and profiled using the Regulatory RNA qPCR Profiler.
Resources
Citations
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- Zhang, J, et al. (2017) Curcumin suppresses cisplatin resistance development partly via modulating extracellular vesicle-mediated transfer of MEG3 and miR-214 in ovarian cancer. Cancer Chemother. Pharmacol..2017 Mar 1; 79(3):479-487. PM ID:28175963
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- Greco, S, et al. (2016) Long noncoding RNA dysregulation in ischemic heart failure. J Transl Med.2016 Jun 18; 14(1):183. PM ID:27317124
- Villamizar, O, et al. (2016) Data in support of transcriptional regulation and function of Fas-antisense long noncoding RNA during human erythropoiesis. Data Brief.2016 Jun 1; 7:1288-95. PM ID:27141526
- Qi, P, Zhou, XY & Du, X. (2016) Circulating long non-coding RNAs in cancer: current status and future perspectives. Mol. Cancer.2016 May 17; 15(1):39. PM ID:27189224
- Villamizar, O, et al. (2016) Data in Brief. Data in Brief.2016 Apr 7; 7:1288–1295. Link:Data in Brief
- Villamizar, O, et al. (2016) Fas-antisense long noncoding RNA is differentially expressed during maturation of human erythrocytes and confers resistance to Fas-mediated cell death. Blood Cells, Molecules, and Diseases.2016 Mar 3; 58:57-66. Link:Blood Cells, Molecules, and Diseases
- Kang, Y, et al. (2016) PCGEM1 stimulates proliferation of osteoarthritic synoviocytes by acting as a sponge for miR-770. J. Orthop. Res..2016 Mar 1; 34(3):412-8. PM ID:26340084
- Thorenoor, N, et al. (2016) Long non-coding RNA ZFAS1 interacts with CDK1 and is involved in p53-dependent cell cycle control and apoptosis in colorectal cancer. Oncotarget.2016 Jan 5; 7(1):622-37. PM ID:26506418
- Patel, K, et al. (2016) LncRNA regulation in mouse lung tumor from dibenzo [def, p] chrysene induced carcinogenesis in a transplacental Nrf2 knockout mouse model. Thesis.;. Link:Thesis
- Carter, G, et al. (2015) Circulating long noncoding RNA GAS5 levels are correlated to prevalence of type 2 diabetes mellitus. BBA Clin.2015 Dec 1; 4:102-7. PM ID:26675493
- Chen, S, et al. (2015) Macrophage infiltration promotes invasiveness of breast cancer cells via activating long non-coding RNA UCA1. Int J Clin Exp Pathol.2015 Oct 14; 8(8):9052-61. PM ID:26464647
- Shen, J, et al. (2015) Exploration of Deregulated Long Non-Coding RNAs in Association with Hepatocarcinogenesis and Survival. Cancers (Basel).2015 Sep 10; 7(3):1847-62. PM ID:26378581
- Shen, J, et al. (2015) Deregulated long non-coding RNAs in hepatocellular carcinoma (HCC). Cancer Research.2015 Aug 1; 75(15S):3818. Link:Cancer Research
- Lin, WC, et al. (2015) Inhibitory effects of high stability fucoxanthin on palmitic acid-induced lipid accumulation in human adipose-derived stem cells through modulation of long non-coding RNA. Food Funct.2015 Jul 1; 6(7):2215-23. PM ID:26057608
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