Cytoplasmic and Nuclear RNA Purification Kit Add to Cart
For the convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues
• Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 45 minutes.
• No genomic DNA contamination in cytoplasmic fraction - Purified cytoplasmic RNA can be used directly in RT-PCR reactions with no DNase treatment required.
• No phenol or chloroform extractions
• Isolate a diversity of RNA species - All nuclear and cytoplasmic RNA species, from large mRNA species down to microRNA (miRNA) and small interfering RNA (siRNA) can be isolated in high quality.
• This kit is available in both 50 prep and 100 prep sizes
Norgen’s Cytoplasmic & Nuclear RNA Purification Kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature,cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA. Norgen’s Cytoplasmic & Nuclear RNA Purification Kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species. The kit is supplied with sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. The isolated high quality RNA is suitable for use in downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array analysis.
Norgen´s Purification Technology
Purification is based on spin column chromatography using Norgen´s proprietary resin as the separation matrix. The cytoplasmic RNA is preferentially purified from the nuclear RNA and other cellular components such as proteins, without the use of phenol or chloroform. The process involves first lysing the cells or tissue of interest with the provided Lysis Solution. The lysate is then separated through centrifugation, with the supernatant containing the cytoplasmic RNA and the pellet containing the nuclear RNA. Binding solution and ethanol is then added to the desired fraction, and the solution is loaded onto a spin-column. Norgen´s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the column, while the contaminating proteins and genomic DNA will be removed in the
flowthrough or retained on the top of the resin. The bound RNA is then washed twice with the provided wash buffer in order to remove any remaining impurities, and the purified RNA is eluted with the elution buffer. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Effective Separation of HeLa Cell Cytoplasmic & Nuclear RNA. Norgen’s Cytoplasmic & Nuclear RNA Purification Kit was used to effectively separate cytoplasmic and nuclear RNA from 0.75 million HeLa cells in triplicate. Panel A: High quality cytoplasmic and nuclear RNA was purified using Norgen´s kit. Note the integrity and quality of both cytoplasmic and nuclear RNA. Ten microliters of each 50 µL elution were run on a 1.5% formaldehyde-agarose gel. Lane M is Norgen´s 1 kb RNA Ladder, lanes 1-3 contain nuclear RNA and lanes 4-6 contain cytoplasmic RNA. Panel B: Ten microliters of the above cytoplasmic and nuclear RNA isolated from HeLa cells using Norgen´s kit was run on a 0.9% agarose DNA gel. Genomic DNA is clearly visible in the nuclear RNA fractions (lanes 1-3), however, no genomic DNA can be detected in the cytoplasmic RNA fractions (lanes 4-6). Note that an optional on-column DNase treatment protocol is provided to remove the genomic DNA in the nuclear fraction.
Genomic DNA-free Cytoplasmic RNA. RT-PCR was performed using human beta actin primers on 2 µL of the 50 µL of cytoplasmic RNA isolated from 1 million HeLa cells using Norgen´s Cytoplasmic & Nuclear RNA Purification Kit. Lane M is Norgen’s PCR Sizer DNA ladder, Lanes 1 and 3 are the negative control (PCR only, without reverse transcript), and Lanes 2 and 4 are the actual RT-PCR that show the expected 166 bp RT-PCR product. The expected amplicon size from the gene copy is the same as that from the RNA transcript. The lack of product in lanes 1 and 3 indicates that no genomic DNA contamination is present in the isolated cytoplasmic RNA. All PCR products were resolved on a 1X TAE, 2% agarose gel.
Specific Separation and Subsequent Amplification of Cytoplasmic and Nuclear Transcripts. Norgen’s Cytoplasmic & Nuclear RNA Purification Kit effectively separates cytoplasmic and nuclear RNA. RT-PCR was performed using human U2 snRNA (left panel) or S14 (right panel) primers on 0.5 µg of either cytoplasmic or nuclear RNA fractions isolated from 1 million HeLa cells using Norgen´s Cytoplasmic & Nuclear RNA Purification Kit. In the left panel, an intense PCR product was observed only in the nuclear fraction when the nuclear-specific U2 snRNA primers were used. In the right panel, the majority of the PCR product for the house-keeping transcript of S14 was observed in the cytoplasmic fraction. The two results combined showed effective separation of the cytoplasmic and nuclear RNA using Norgen’s Cytoplasmic & Nuclear RNA Purification Kit. All PCR products were resolved on a 1X TAE, 1.5% agarose DNA gel.
- Kouwaki T, Fukushima Y, Daito T, Sanada T, Yamamoto N, Mifsud EJ, Leong CR, Tsukiyama-Kohara K, Kohara M, Matsumoto M, Seya T, Oshiumi H. (2016) Extracellular Vesicles Including Exosomes Regulate Innate Immune Responses to Hepatitis B Virus Infection. Frontiers in Immunology
- Kishikawa T, Otsuka M,, Yoshikawa T, Ohno M, Ijichi H, Koike K. (2016) Satellite RNAs promote pancreatic oncogenic processes via the dysfunction of YBX1. Nature Communications
- Derek A. Oldridge, Andrew C. Wood, Nina Weichert-Leahey, Ian Crimmins, Robyn Sussman, Cynthia Winter, Lee D. McDaniel, Maura Diamond, Lori S. Hart, Shizhen Zhu, Adam D. Durbin, Brian J. Abraham, Lars Anders, Lifeng Tian, Shile Zhang, Jun S. Wei, Javed Khan, Kelli Bramlett, Nazneen Rahman, Mario Capasso, Achille Iolascon, Daniela S. Gerhard, Jaime M. Guidry Auvil, Richard A. Young, Hakon Hakonarson et al. (2015) Genetic predisposition to neuroblastoma mediated by a LMO1 super-enhancer polymorphism. Nature
- He C, Ding JW, Li S, Wu H, Jiang YR, Yang W, Teng L, Yang J, Yang J. (2015) The Role of Long Intergenic Noncoding RNA p21 in Vascular Endothelial Cells. DNA and Cell Biology
- Kim RH, Kang MK, Kim T, Yang P, Bae S, Williams DW, Phung S, Shin KH, Hong C, Park NH. (2015) Regulation of p53 during senescence in normal human keratinocytes. Aging Cell
- Cao S, Moss W, O'Grady T, Concha M, Strong MJ, Wang X, Yu Y, Baddoo M, Zhang K, Fewell C, Lin Z, Dong Y, Flemington EK. (2015) New non-coding lytic transcripts derived from the Epstein Barr virus latency origin of replication oriP are hyper-edited, bind the paraspeckle protein, NONO/p54nrb, and support lytic viral transcription. Journal of Virology
- Inma Gonzalez, Roberto Munita, Eneritz Agirre, Travis A Dittmer, Katia Gysling, Tom Misteli & Reini F Luco. (2015) A lncRNA regulates alternative splicing via establishment of a splicing-specific chromatin signature. Nature Structural & Molecular Biology
- Rozanne Arulanandam, Cory Batenchuk, Oliver Varette, Chadi Zakaria, Vanessa Garcia, Nicole E. Forbes, Colin Davis, Ramya Krishnan, Raunak Karmacharya, Julie Cox, Anisha Sinha, Andrew Babawy, KatherineWaite, EricaWeinstein, Theresa Falls, Andrew Chen, Jeff Hamill, Naomi De Silva, David P. Conrad, Harold Atkins, Kenneth Garson, Carolina Ilkow, Mads Kærn, Barbara Vanderhyden, Nahum Sonenberg, Tommy Alain, Fabrice Le Boeuf, John C. Bell1, & Jean-Simon Diallo. (2015) Microtubule disruption synergizes with oncolytic virotherapy by inhibiting interferon translation and potentiating bystander killing. Nature Communications
- Xu TP, Liu XX, Xia R, Yin L, Kong R, Chen WM, Huang MD, Shu YQ. (2015) SP1-induced upregulation of the long noncoding RNA TINCR regulates cell proliferation and apoptosis by affecting KLF2 mRNA stability in gastric cancer. Oncogene
- Fujun Yu, Jianjian Zheng, Yuqing Mao, Peihong Dong, Zhongqiu Lu, Guojun Li, Chuanyong Guo, Zhanju Liu and Xiaoming Fan. (2015) Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA. The Journal of Biological Chemistry
- Sakaguchi M, Watanabe M, Kinoshita R, Kaku H, Ueki H, Futami J, Murata H, Inoue Y, Li SA, Huang P, Putranto EW, Ruma IM, Nasu Y, Kumon H, Huh NH. (2014) Dramatic Increase in Expression of a Transgene by Insertion of Promoters Downstream of the Cargo Gene. Molecular Biotechnology
- Andrew E Jaffe, Jooheon Shin, Leonardo Collado-Torres, Jeffrey T Leek, Ran Tao, Chao Li, Yuan Gao, Yankai Jia, Brady J Maher, Thomas M Hyde, Joel E Kleinman & Daniel R Weinberger. (2014) Developmental regulation of human cortex transcription and its clinical relevance at single base resolution. Nature Neuroscience
- Cao S, Strong MJ, Wang X, Moss WN, Concha M, Lin Z, O'Grady T, Baddoo M, Fewell C, Renne R, Flemington EK. (2014) High-Throughput RNA sequencing based virome analysis of 50 lymphoma cell lines from the Cancer Cell Line Encyclopedia project. Journal of Virology
- Ning-xia Sun, Chen Ye, Qian Zhao, Qing Zhang, Chen Xu, Shao-bing Wang, Zhi-jun Jin, Shu-han Sun, Fang Wang, Wen Li mail. (2014) Long Noncoding RNA-EBIC Promotes Tumor Cell Invasion by Binding to EZH2 and Repressing E-Cadherin in Cervical Cancer. PLOS ONE
- Zaghlool A, Ameur A, Nyberg L, Halvardson J, Grabherr M, Cavelier L, Feuk L. (2013) Efficient cellular fractionation improves RNA sequencing analysis of mature and nascent transcripts from human tissues. BMC Biotechnology
- Li J, Chen Y, Qin X, Wen J, Ding H, Xia W, Li S, Su X, Wang W, Li H, Zhao Q, Fang T, Qu L, Shao N. (2013) MiR-138 downregulates miRNA processing in HeLa cells by targeting RMND5A and decreasing Exportin-5 stability. Nucleic Acids Research
- Lee H, Komano J, Saitoh Y, Yamaoka S, Kozaki T, Misawa T, Takahama M, Satoh T, Takeuchi O, Yamamoto N, Matsuura Y, Saitoh T, Akira S. (2013) Zinc-finger antiviral protein mediates retinoic acid inducible gene I-like receptor-independent antiviral response to murine leukemia virus. Proceedings of the National Academy of Sciences of the United States of America
- Varia S, Potabathula D, Deng Z, Bubulya A, Bubulya PA. (2013) Btf and TRAP150 have distinct roles in regulating subcellular mRNA distribution. Nucleus
- Spadaro F, Cecchetti S, Purificato C, Sabbatucci M, Podo F, Ramoni C, Gessani S, Fantuzzi L. (2013) Nuclear Phosphoinositide-Specific Phospholipase C β1 Controls Cytoplasmic CCL2 mRNA Levels in HIV-1 gp120-Stimulated Primary Human Macrophages. PLoS One
- Jeck WR, Sorrentino JA, Wang K, Slevin MK, Burd CE, Liu J, Marzluff WF, Sharpless NE. (2013) Circular RNAs are abundant, conserved, and associated with ALU repeats. RNA: A Publication of the RNA Society
- Liua Y, Liua H, Zoua J, Zhanga B and Yuana Z. (2013) Dengue virus subgenomic RNA induces apoptosis through the Bcl-2-mediated PI3k/Akt signaling pathway. Virology
- Gumireddy K, Li A, Yan J, Setoyama T, Johannes GJ, Orom UA, Tchou J, Liu Q, Zhang L, Speicher DW, Calin GA, Huang Q. (2013) Identification of a long non-coding RNA-associated RNP complex regulating metastasis at the translational step. The EMBO Journal
- Wan G,Zhang X, Langley R, Liu Y, Hu X, Han C, Peng G, Ellis L, Jones S, Lu X. (2013) DNA-Damage-Induced Nuclear Export of Precursor MicroRNAs Is Regulated by the ATM-AKT Pathway. Cell
- Meyer JL, Huber JA. (2013) Strain-level genomic variation in natural populations of Lebetimonas from an erupting deep-sea volcano. The IMSE Journal
- Carrieri C, Cimatti L, Biagioli M, Beugnet A, Zucchelli S, Fedele S, Pesce E, Ferrer I, Collavin L, Santoro C, Forrest AR, Carninci P, Biffo S, Stupka E, Gustincich S. (2012) Long non-coding antisense RNA controls Uchl1 translation through an embedded SINEB2 repeat. Nature
- Iglesias-Platas I, Martin-Trujillo A, Cirillo D, Court F, Guillaumet-Adkins A, Camprubi C, Bourc'his D, Hata K, Feil R, Tartaglia G, Arnaud P, Monk D. (2012) Characterization of Novel Paternal ncRNAs at the Plagl1 Locus, Including Hymai, Predicted to Interact with Regulators of Active Chromatin. PLoS One
- Li N, Zhang L, Chen L, Feng W, Xu Y, Chen F, Liu X, Chen Z, Liu W. (2012) MxA inhibits hepatitis B virus replication by interaction with core protein HBcAg. Hepatology
- Chiappinelli KB, Haynes BC, Brent MR, Goodfellow PJ. (2012) Reduced DICER1 elicits an interferon response in endometrial cancer cells. Molecular Cancer Research
- Walker EJ, Zhang C, Castelo-Branco P, Hawkins C, Wilson W, Zhukova N, Alon N, Novokmet A, Baskin B, Ray P, Knobbe C, Dirks P, Taylor MD, Croul S, Malkin D, Tabori U. (2011) Monoallelic Expression Determines Oncogenic Progression and Outcome in Benign and Malignant Brain Tumors. Cancer Research
- Concha M, Wang X, Cao S, Baddoo M, Fewell C, Lin Z, Hulme W, Hedges D, McBride J, Flemington EK. (2011) Identification of new viral genes and transcript isoforms during Epstein-Barr virus reactivation using RNA-seq. Journal of Virology
- Fan YH, Nadar M, Chen CC, Weng CC, Lin YT, Chang RY. (2011) Small Noncoding RNA Modulates Japanese Encephalitis Virus Replication and Translation in trans. Virology Journal
- Jin X, Sun T, Zhao C, Zheng Y, Zhang Y, Cai W, He Q, Taira K, Zhang L, Zhou D. (2011) Strand antagonism in RNAi: an explanation of differences in potency between intracellularly expressed siRNA and shRNA. Nucleic Acids Resarch
- Jeffries CD, Fried HM, Perkins DO. (2011) Nuclear and cytoplasmic localization of neural stem cell microRNAs. RNA
- Melo SA, Moutinho C, Ropero S, Calin GA, Rossi S, Spizzo R, Fernandez AF, Davalos V, Villanueva A, Montoya G, Yamamoto H, Schwartz S Jr, Esteller M. (2010) A genetic defect in exportin-5 traps precursor microRNAs in the nucleus of cancer cells. Cancer Cell
- Yap KL, Li S, Muñoz-Cabello AM, Raguz S, Zeng L, Mujtaba S, Gil J, Walsh MJ, Zhou MM. (2010) Molecular Interplay of the Noncoding RNA ANRIL and Methylated Histone H3 Lysine 27 by Polycomb CBX7 in Transcriptional Silencing of INK4a. Mol Cell
- Berulava T, Horsthemke B. (2010) The obesity-associated SNPs in intron 1 of the FTO gene affect primary transcript levels. Eur J Hum Genet
- Bernot D, Barruet E, Poggi M, Bonardo B, Alessi MC, Peiretti F. (2010) Down-regulation of tissue inhibitor of metalloproteinase-3 (TIMP-3) expression is necessary for adipocyte differentiation. Journal of Biological Chemistry
- Jeffries CD, Perkins DO, Guan X. (2010) Gene processing control loops suggested by sequencing, splicing, and RNA folding. BMC Bioformatics
- Zhang X, Chen S, Yoo S, Chakrabarti S, Zhang T, Ke T, Oberti C, Yong SL, Fang F, Li L, de la Fuente R, Wang L, Chen Q. (2008) Mutation in Nuclear Pore Component NUP155 Leads to Atrial Fibrillation and Early Sudden Cardiac Death. Cell
- Chen N, Onisko B, Napoli J. (2008) The Nuclear Transcription Factor RARa Associates with Neuronal RNA Granules and Suppresses Translation. Journal of Biological Chemistry