Cytoplasmic and Nuclear RNA Purification Kit For the convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Cytoplasmic and Nuclear RNA Purification Kit For the convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues USD$385.00 SKU 21000 Format Spin Column Size 50 Preps 100 Preps Quantity Overview Details Documentation Citations Overview Excellent separation and purification of cytoplasmic and nuclear RNA Convenient and fast spin column format High quality and yield of RNA Isolate full diversity of RNA (including microRNA) without phenol Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR 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 both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size. Background 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. Details Supporting Data Previous Figure 1. 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. Figure 2. Superior Separation of HeLa Cell Cytoplasmic & Nuclear RNA. Norgen's Cytoplasmic & Nuclear RNA Purification Kit provides better separation of cytoplasmic and nuclear RNA from 0.8 million HeLa cells when compared to a leading competitor’s product. Panel A: Cytoplasmic and nuclear RNA purified from HeLa cells using Norgen's kit and a competitor's kit. Ten microliters of each 50 µL elution (Norgen or competitor's kit) of the cytoplasmic (C) or nuclear (N) RNA were run on a 1.5% formaldehyde-agarose gel. Higher yields of RNA with good integrity were isolated using Norgen's kit. Panel B: Ten microliters of the cytoplasmic and nuclear RNA isolated from HeLa cells using Norgen's kit and the competitor's kit was run on a 0.9% agarose gel. Genomic DNA only co-migrates with the nuclear fraction in RNA isolated using Norgen’s Cytoplasmic & Nuclear RNA Purification Kit, not the cytoplasmic fraction. Note that an optional on-column DNase treatment protocol is provided to remove the genomic DNA in the nuclear fraction. In contrast, significant genomic DNA contamination was observed in the cytoplasmic fraction of the RNA isolated using the competitor's kit. Figure 3. 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. Figure 4. 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 (Panel A) or S14 (Panel B) 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 Panel A, an intense PCR product was observed only in the nuclear fraction when the nuclear-specific U2 snRNA primers were used. In Panel B, 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. Next Click for expanded view Kit Specifications Maximum Column Binding Capacity Up to 50 µg RNA Maximum Column Loading Volume 650 µL Size of RNA Purified All sizes, including small RNA (< 200 nt) Time to Complete 10 Purifications 45 minutes RNA Yield HeLa (1 x 106) - Cytoplasmic RNA HeLa (1 x 106) - Nuclear RNA 15 µg ≤ 3.5 µg Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. Component Cat. 21000 (50 preps) Cat. 37400 (100 preps) Lysis Buffer J 20 mL 2 x 20 mL Buffer SK 40 mL 2 x 40 mL Wash Solution A 38 mL 2 x 38 mL Elution Buffer E 6 mL 2 x 6 mL Mini Spin Columns 50 100 Collection Tubes 50 100 Elution Tubes (1.7 mL) 50 100 Product Insert 1 1 Documentation PROTOCOLS (21000) Cytoplasmic & Nuclear RNA Purification Kit - Protocol (50 prep) (37400) Cytoplasmic & Nuclear RNA Purification Kit - Protocol (100 prep) APPLICATION NOTES Isolation of Fractionated RNA From Hela Cells Using Norgen's Cytoplasmic Nuclear RNA Purification Kit SAFETY DATA SHEETS (21000, 37400) Cytoplasmic & Nuclear RNA Purification Kit - Safety Data Sheet POSTERS Cytoplasmic and Nuclear RNA Purification Kit Effects of The Adenovirus Type 5 Tripartite Leader Sequence With Partial and Comple RELATED BLOGS 3 Reasons why Norgen’s Industry-Leading Patented RNA Purification Technology is Best-in-Class Citations Powered by Bioz See more details on Bioz Related Products Leukocyte RNA Purification Kit microRNA Purification Kit RNase-Free DNase I Kit Total RNA Purification Kit
Cytoplasmic and Nuclear RNA Purification Kit For the convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Cytoplasmic and Nuclear RNA Purification Kit For the convenient purification of cytoplasmic and nuclear RNA from cultured cells and tissues USD$385.00 SKU 21000 Format Spin Column Size 50 Preps 100 Preps Quantity