microRNA Purification Kit For the rapid purification of microRNA without phenol For research use only and NOT intended for in vitro diagnostics. Powered by Bioz microRNA Purification Kit For the rapid purification of microRNA without phenol Price USD$249.00 SKU 21300 Size 25 Preps Quantity Overview Details Documentation Citations Overview Isolate and extract high quality & quantity miRNA in 30 minutes Two column kit- eliminate large RNA on the first column and capture microRNA on a second column Elute microRNA in 25 µL ready for miRNA profiling Process a wide range of samples- cell, tissue, bacteria, bodily fluids, etc. Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood. Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water. The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications. Background These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation. Details Supporting Data Previous Figure 1. High Quality of Isolated RNA with Better Size Exclusion. Small RNA was isolated from HeLa cells using Norgen's microRNA Purification Kit (Lanes C and D) and a competitor's kit (Lanes A and B). Ten microliters of the 50 µL purified small RNA were run on an 8% urea-agarose gel. Lane M is Norgen's 100b RNA ladder. Note that Norgen's kit is isolating only the small (<200 nt) RNA species, with no contaminating larger RNA. Figure 2. Efficient Removal of Large RNA Species. Different RNA species were isolated from 106 HeLa cells, resolved on an Agilent® Lab-On-A-Chip, and electropherograms were generated. Panel A contains all the RNA species present in 106 HeLa cells as isolated with Norgen's Total RNA Purification Kit and acts as a control. Panel B and C contain RNA that was isolated from 106 HeLa cells using Norgen’s microRNA Purification Kit. One microliter of the 50 µL purified RNA for each fraction was loaded. Panel B shows the large RNA species removed using the Large RNA Removal Columns, and no small RNA can be detected. Panel C shows the small RNA that is isolated using the microRNA Enrichment Columns and shows that there is no contamination of the small RNA with any large RNA species above 200 nt. This demonstrates the effective separation of the small RNA from the large RNA species using Norgen's microRNA Purification Kit. Figure 3. Effective Small RNA Amplification by End-Point RT-PCR. Norgen's microRNA Purification Kit isolates small RNA that could be amplified in RT-PCR using the poly (A) polymerase extension method. Small RNA was isolated from 1 million HeLa cells, and 10 µL of the 20 µL isolated small RNA were polyadenylated in a 50 µL Poly-(A)-Polymerase reaction. Seven microliters of the polyadenylated RNA were used in a 20 µL reverse transcription reaction with a poly T adaptor primer. One microliter of the reverse transcription was used in a 20 µL PCR reaction with primers against the human microRNAs (miR-21) and 5S rRNA. Panel A shows the amplification of the miR-21 transcript from small RNAs while Panel B shows the 5S rRNA amplification from small RNAs. Lane 1 in both panels shows the results when total RNA isolated from 1 million HeLa cells using Norgen's Total RNA Purification Kit was used as a control. Lanes 2 and 3 contain the successful RT-PCR when the microRNA isolated using Norgen's microRNA Purification Kit was used as the template, and Lanes 4 contain the non-template control. The RT-PCR was successful for both reactions using the microRNA as the template. All PCR products were resolved on a 1X TAE, 3% agarose gel using Norgen's PCR Ranger as the molecular weight ladder. Figure 4. Better Recovery of miRNAs by Norgen's microRNA Purification Kit. Norgen's microRNA Purification Kit recovers microRNA more effectively than its competitors. Small RNA was isolated from 0.75 million HeLa cells using Norgen's microRNA Purification Kit and a competitors' kits. Relative expression of (A) miR-21, (B) miR-19, and (C) 5S rRNA was determined by RT-qPCR of polyadenylated total RNA samples. RT-qPCR was performed according to Shi and Chiang (2005). Fifteen microliters of the 50 µL isolated RNA were polyadenylated in a 50 µL Poly-(A)-Polymerase reaction. Four microliters of the polyadenylated RNA were used in a 20 µL reverse transcription reaction with a poly T adaptor primer. One microliter of the reverse transcription was then used in a 20 µL qPCR reaction with primers against the human microRNAs (miR-19 and miR-21) and house-keeping small RNA (5S rRNA). The resulting threshold cycle (Ct) values were summarized in the graph. Blue = Norgen's microRNA Purification Kit; orange = Silica-based Competitor micro-RNA Kit; green = Silica-based Competitor Total RNA Kit; red = Phenol-based reagent protocol; yellow = No Template Control. Norgen's microRNA Purification Kit recovered more miRNAs (lower Ct) than the competitor's microRNA-specific Kit. Next Click for expanded view Kit Specifications Maximum Column Binding Capacity Up to 50 μg RNA Maximum Column Loading Volume 650 μL Minimum Elution Volume 20 μL Size of RNA Purified All sizes, including small RNA < 200 nt Time to Complete 10 Purifications 25 minutes Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood 3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment. Component Cat. 21300 (25 preps) Buffer RL 40 mL Wash Solution A 38 mL Elution Solution A 6 mL Large RNA Removal Column 25 microRNA Enrichment Column 25 Collection Tube 50 Elution Tubes (1.7 mL) 50 Product Insert 1 Documentation PROTOCOLS (21300) microRNA Purification Kit - Protocol (25 prep) APPLICATION NOTES Non-Organic-Based Isolation of Mammalian microRNA using Norgen’s microRNA Purification Kit Effect of RNA Isolation Method on microRNA Quantity and Quality in Plasma: A Comparative Study SAFETY DATA SHEETS (21300) microRNA Purification Kit - Safety Data Sheet POSTERS Silicon Carbide As A Novel RNA Affinity Medium With Improved Sensitivity and Size Diversity - microRNA 2008 FLYERS Flyer RELATED BLOGS 3 Reasons why Norgen’s Industry-Leading Patented RNA Purification Technology is Best-in-Class How to Reduce False Negative SARS-CoV-2 Results with Silicon Carbide Citations Title Nrf2 Plays a Key Role in Erythropoiesis during Aging Journal Antioxidants. 2024. Authors Serge Cedrick Toya Mbiandjeu, Angela Siciliano, Alessandro Mattè, Enrica Federti, Massimiliano Perduca, Davide Melisi, Immacolata Andolfo, Angela Amoresano, Achille Iolascon, Maria Teresa Valenti, Francesco Turrini, Michele Bovi, Arianna Pisani, Antonio Recchiuti, Domenico Mattoscio, Veronica Riccardi,Luca Dalle Carbonare, Carlo Brugnara, Narla Mohandas, and Lucia De Franceschi Title Early Salivary miRNA Expression in Extreme Low Gestational Age Newborns Journal Life. 2022. Authors Roopa Siddaiah, Lucy Emery, Heather Stephens, Ann Donnelly, Jennifer Erkinger, Kimberly Wisecup, Steven D. Hicks, Yuka Imamura Kawasawa, Christiana Oji-Mmuo, Shaili Amatya and Patricia Silveyra Title MicorRNA-195 links long non-coding RNA SEMA3B antisense RNA 1 (head to head) and cyclin D1 to regulate the proliferation of glioblastoma cells Journal Bioengineered. 2022. Authors Kaijun Liu, Yan Deng, Yongxia Yang, Hui Wang & Ping Zhou Title Cancer cells exploit an orphan RNA to drive metastatic progression Journal Nature Medicine. 2018. Authors Lisa Fish, Steven Zhang, Johnny X. Yu, Bruce Culbertson, Alicia Y. Zhou, Andrei Goga & Hani Goodarzi Title Expression of miRNA-140 in Chondrocytes and Synovial Fluid of Knee Joints in Patients with Osteoarthritis Journal Chinese Medical Sciences Journal. 2016. Authors Hai-Bo S, Yi Z, Zong-Ke Z, Fu-Xing P, Yan-Rong L, Jing-Qiu C, Bin S. Title Genomewide Association Study of African Children Identifies Association of SCHIP1 and PDE8A with Facial Size and Shape Journal PLOS One. 2016. Authors Cole JB, Manyama M, Kimwaga E, Mathayo J, Larson JR, Liberton DK, Lukowiak K, Ferrara TM, Riccardi SL, Li M, Mio W, Prochazkova M,, Williams T, Li H, Jones KL, Klein OD, Santorico SA, Hallgrimsson B, Spritz RA Title Methods of Detecting Lung Cancer Journal United States Patent Application 20160108478. 2016. Authors Delfour, Olivier, Michot, Bernard Title The human PMR1 endonuclease stimulates cell motility by down regulating miR-200 family microRNAs Journal Nucleic Acid Research. 2016. Authors Gu SQ, Gallego-Perez D, McClory SP, Shi J, Han J, Lee LJ, Schoenberg DR Title Analysis of hepatic microRNA alterations in response to hepatitis B virus infection and pegylated interferon alpha-2a treatment Journal Experimental Biology and Medicine. 2016. Authors Jinato T, Chuaypen N, Poomipak W, Praianantathavorn K, Makkoch J, Kiatbumrung R, Jampoka K, Tangkijvanich P, Payungporn S Title MEMO1 drives cranial endochondral ossification and palatogenesis Journal Developmental Biology. 2015. Authors Van Otterloo E, Feng W, Jones KL, Hynes NE, Clouthier DE, Niswander L, Williams T Title Human microRNAs profiling in response to influenza A viruses (subtypes pH1N1, H3N2, and H5N1) Journal Experimental Biology and Medicine. 2015. Authors Makkoch J, Poomipak W, Saengchoowong S, Khongnomnan K, Praianantathavorn K, Jinato T, Poovorawan Y, Payungporn S Title Circulating microRNA Profiling Needs Further Refinement Before Clinical Use in Patients With Aortic Stenosis Journal Valvular Heart Disease. 2015. Authors Sean Coffey, MBBS, Michael J. A. Williams, MD, L. Vicky Phillips, Gregory T. Jones, PhD Title Identification of MicroRNAs and Transcript Targets in Camelina sativa by Deep Sequencing and Computational Methods Journal PLoS ONE. 2015. Authors Saroj Poudel, Niranjan Aryal, Chaofu Lu Title MicroRNA-181b regulates ALX/FPR2 expression and proresolution signaling in human macrophages Journal The Journal of Biological Chemistry. 2014. Authors Anna Maria Pierdomenico, Antonio Recchiuti, Felice Simiele, Marilina Codagnone, Veronica Cecilia Mari, Giovanni Davi and Mario Romano Title Role of TLR13 in innate immune recognition of group B streptococci Journal Infection and Immunity. 2014. Authors Giacomo Signorino, Nastaran Mohammadi, Francesco Patanè, Marco Buscetta, Mario Venza, Isabella Venza, Giuseppe Mancuso, Angelina Midiri, Lena Alexopoulou, Giuseppe Teti, Carmelo Biondo and Concetta Beninati. Title Quantification of Circulating miRNAs in Plasma: Effect of Preanalytical and Analytical Parameters on Their Isolation and Stability. Journal The Journal of Molecular Diagnostics. 2013. Authors Sourvinou IS, Markou A, Lianidou ES. Title A Panel of four MicroRNAs accurately differentiates Malignant from Benign Indeterminate Thyroid Lesions on Fine Needle Aspiration. Journal Clinical Cancer Research. 2012. Authors Xavier M. Keutgen, Filippo Filicori, Michael J. Crowley, et al. Title Effective inhibition of replication of infectious bursal disease virus by miRNAs delivered by vectors and targeting the VP2 gene Journal Journal of Virological Methods.65(2):127-32. 2010. Authors Wang Y, Sun H, Shen P, Zhang X, Xia X, Xia B. Title Bacterial recognition by TLR7 in the lysosomes of conventional dendritic cells Journal Nature Immunology.10,587 - 594. 2009. Authors Mancuso G,Gambuzza M,Midiri A,Biondo C,Papasergi S,Akira S,Teti G, Beninati C. Title Novel expression of microRNAs in serum samples of Iraqi breast cancer women Journal American Journal of BioMedicine. 2014. Authors Zaynab Saad; Muhammed Arif; Nahi Yassen; Hameed Jasim; Majed Jelawe; James Brown Title Expanding the annotation of zebrafish microRNAs based on small RNA sequencing Journal Gene. 2014. Authors Thomas Desvignes, Michael J. Beam, Peter Batzel, Jason Sydes, John H. Postlethwait Title AcvR1-mediated BMP signaling in second heart field is required for arterial pole development: Implications for myocardial differentiation and regional identity. Journal Developmental Biology. 2014. Authors Thomas PS, Rajderkar S, Lane J, Mishina Y, Kaartinen V. Title Effective inhibition of infectious bursal disease virus replication by 1recombinant avian adeno-associated virus-delivered microRNAs. Journal Journal of Genetic Virology. 90: 1417-1422. Authors Wang Y, Sun H, Shen P, Zhang X, Xiaoli X. Related Services NGS RNA Isolation Related Products Cytoplasmic & Nuclear RNA Purification Kit Leukocyte RNA Purification Kit RNA/DNA/Protein Purification Plus Kit Total RNA Purification Kit
microRNA Purification Kit For the rapid purification of microRNA without phenol For research use only and NOT intended for in vitro diagnostics. Powered by Bioz microRNA Purification Kit For the rapid purification of microRNA without phenol Price USD$249.00 SKU 21300 Size 25 Preps Quantity