RNA Clean-Up and Concentration Kits For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol For research use only and NOT intended for in vitro diagnostics. Powered by Bioz RNA Clean-Up and Concentration Kits For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol SKU 23600 Format Spin Column High Throughput Micro-Elute Size 50 Preps 100 Preps Contact Distributor Request sample Overview Details Documentation Citations Overview Clean-up & concentrate total RNA (including miRNA) in minutes Clean-up & concentrate RNA from TRIzol®, TRI Reagent®, etc Clean-up RNA from contaminants including enzymes, primers, nucleotides Rapid spin-column protocol, elute in 20 µL 96-well format for high throughput processing & Micro-Elute spin column formats for 8 µL are available Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix These kits are designed to clean-up and concentrate RNA (including miRNA) from TRIzol® and TRI Reagent®, enzymatic reactions, in vitro transcription, labelling reactions, etc. These are robust kits for all clean-up and concentration purposes for up to 35 µg of RNA in solution. The purified RNA is of the highest purity and integrity, and can be used in a number of downstream applications. These kits purify all sizes of RNA, from large mRNA and ribosomal RNA down to miRNA, siRNA and lncRNA. RNA Clean-Up and Concentration Kit (Spin Column) Norgen’s RNA Clean-Up and Concentration Kit provides a rapid method for the purification, cleanup and concentration of up to 35 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 20 µL. Complete 10 purifications in 20 minutes. RNA Clean-Up and Concentration 96-Well Kit (High Throughput) Norgen’s RNA Clean-Up and Concentration 96-Well Kit provides a rapid method for the purification, cleanup and concentration of up to 50 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 75 µL. Complete 10 purifications in 30 minutes. RNA Clean-Up and Concentration Micro-Elute Kit (Micro-Elute) Norgen’s RNA Clean-Up and Concentration Micro-Elution Kit provides a rapid method for the purification, cleanup and concentration of up to 10 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. This kit is made for eluting RNA in smaller volumes of 8 µL for all types of downstream applications, for the highest concentration of RNA sample. Complete 10 purifications in 20 minutes. Protocol Details Supporting Data Previous Figure 1. Effective Clean-Up to Produce High Quality Total RNA with Complete Size Diversity. Norgen's RNA Clean-Up and Concentration Kit effectively cleans up RNA isolated from phenol-based extractions without the loss of RNA diversity by retaining all RNA species including small RNAs. Total RNA was isolated from 5 x 108 E. coli using a competitor's phenol-based RNA extraction reagent. The resulting RNA was then purified using Norgen’s RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Norgen's Total RNA Purification Kit (#17200, no phenol required) and the phenol-based RNA extraction reagent only without any clean-up. Resolution of 7 µL of the 50 µL purified RNA on a 1X MOPS, 1.5% formaldehyde-agarose gel showed the RNA extracted with phenol was successfully cleaned-up by Norgen's RNA Clean-Up and Concentration Kit without the loss of small RNA species. Figure 2. Effective Clean-Up to Produce High Quality Total RNA Compatible to Bioanalyzer Analysis. Total RNA was isolated from 0.75 million HeLa cells using a competitor's phenol-based RNA extraction reagent. The resulting RNA was then purified using Norgen's RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Norgen's Total RNA Purification Kit (#17200, no phenol required) or the phenol-based RNA extraction reagent only without clean-up. Resolution of 1 µL of the 50 µL purified RNA on an Agilent RNA Nano 6000 Chip showed that without clean-up, the RNA sample isolated using the phenol-based RNA extraction reagent only resolved poorly on a bioanalyzer. In contrast, RNA extracted with the phenol-based RNA extraction reagent followed by clean-up by Norgen's RNA Clean-Up and Concentration Kit resolved properly on the bioanalyzer showing excellent quality. Figure 3. Effective Concentration and Detection of Low Amounts of RNA Input. Norgen's RNA Clean-Up and Concentration Kit can effectively concentration RNA inputs in the picogram range. Increasing amounts of HeLa RNA in 50 µL input volumes were concentrated to 20 µL using Norgen's RNA Clean-Up and Concentration Kit. A 10 µL aliquot of the purified RNA was then used as the template in a qRT-PCR reaction to detect the S14 gene. The amounts indicated on the graph correspond to the amount of RNA that was used as the input for the qRT-PCR reaction, demonstrating the consistent performance of the kit even in the picogram range. Figure 4. Effective Clean-Up to Produce High Quality RNA Transcripts. Norgen's RNA Clean-Up and Concentration Kit effectively cleans up RNA transcripts with high recovery. Three RNA transcripts (1-3) of different sizes (266n, 161n and 419n, respectively) were produced using Norgen's RNA Clean-Up and Concentration Kit. The resulting RNA transcripts were then purified using Norgen's RNA Clean-Up and Concentration Kit. Resolution of 7.5 µL of the 50 µL cleaned RNA (C) as well as an equal volume of the uncleaned input (I) on a 1X MOPS, 1.5% formaldehyde-agarose gel showed the RNA transcript was successfully cleaned-up by Norgen's RNA Clean-Up and Concentration Kit. Figure 5. Effective Clean-Up to Produce High Quality RNA Transcripts Compatible to Bioanalyzer Analysis. Norgen's RNA Clean-Up and Concentration Kit effectively cleans up RNA transcripts with high recovery. Three RNA transcripts (1-3) of different sizes (266n, 161n and 419n, respectively) were produced using Norgen's RNA Clean-Up and Concentration Kit. Resolution of 1 µL of the 50 µL cleaned RNA transcripts on an Agilent RNA Nano 6000 Chip showed that RNA transcripts cleaned by Norgen's RNA Clean-Up and Concentration Kit resolved properly on the bioanalyzer showing excellent quality. Figure 6. Excellent Quality of Concentrated RNA. Total RNA isolated from HeLa cells (2 µg) was concentrated to 8 µL using the RNA Clean-Up and Concentration Micro-Elute Kit. The excellent quality is indicated by the electropherogram generated using the Agilent 2100 Bioanalyzer (RIN > 9). The concentrated RNA is a true 'total RNA' as can be observed by the presence of small RNA species. Figure 7. High Recovery of Concentrated RNA. Total RNA isolated from HeLa cells (17.5 µL) was concentrated to 8 µL using the RNA Clean-Up and Concentration Micro-Elute Kit resulting in a 2-2.2 fold increase in RNA concentration, as was measured by Agilent 2100 Bioanalyzer quantification. Figure 8. Concentration of Total RNA. The indicated amounts of HeLa RNA were concentrated using the RNA Clean-Up and Concentration Micro-Elute Kit. For each input amount, a 17.5 µL volume was processed and the RNA eluted in 8 µL, resulting in a 2.2 fold concentration. Three microliters of each eluate were used in 10 µL RT reactions with the oligo-dT primer, followed by real-time PCR using 3 µL of the resulting cDNA and primers specific for the human RPS15 gene. On average, amplifications of the concentrated RNA reached threshold 1 Ct value before the input RNA samples, which was expected based on the concentration factor. The lowest input amount used (240 pg) was only amplified when concentrated (most right sample in figure). Figure 9. Concentration of miRNA. The indicated amounts of HeLa RNA were concentrated using the RNA Clean-Up and Concentration Micro-Elute Kit. Three microliters of each 8 µL eluate were used in 10 µL RT reactions with the miR-21-SLR primer, followed by real-time PCR using 3 µL of the resulting cDNA and the forward primer specific for miR-21. Figure 10. Concentration of RNA prior to Next Generation Sequencing (NGS) applications. Total RNA was purified from 200 µL of plasma collected on EDTA blood tubes using Norgen's Total RNA Purification Kit (Cat. 17200) and eluted in 50 µL of elution solution. The same RNA was also concentrated two-fold using the Micro-Elute RNA Column by eluting in 25 µL of elution solution. Five microliters of both the RNA without additional concentration and the 2X concentrated RNA were used as inputs to generate RNA libraries (using the NEBNext® Small RNA Library Prep Set for Illumina® and following manufacturer’s instructions) for small RNA NGS on the MiSeq (Illumina) platform. A) The prepared small RNA libraries were visualized on a 6% TBE polyacrylamide gel, where the library prepared with 2X concentrated RNA contained more ligated/indexed miRNA cDNA (147-160 bp) products than the library prepared using the RNA without concentration. B) The cDNA was extracted from excised gel bands and interrogated using the Agilent 2100 Bioanalyzer (High Sensitivity DNA Assay). As would be expected based on input, the small RNA library prepared with the 2X concentrated RNA sample was approximately two times more concentrated than the library prepared with RNA without prior concentration (39.8 vs 21.2 nM, respectively). Next Click for expanded view Kit Specifications (Spin Column) Maximum Column Binding Capacity 35 μg Size of RNA Purified All sizes, including small RNA (<200 nt) Maximum Amount of Starting Material 35 μg of RNA Minimum Elution Volume 20 μL Time to Complete 10 Purifications 20 minutes Average Recovery ≥ 90% 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. 23600 (50 preps) Cat. 43200 (100 preps) Cat. 25100 (192 preps) Cat. 61000 (50 preps) Buffer RL 40 mL 2 x 40 mL 2 x 40 mL 40 mL Wash Solution A 38 mL 2 x 38 mL 2 x 38 mL 38 mL Elution Solution A 6 mL 2 x 6 mL 2 x 20 mL 6 mL Column Activation Solution - - - 30 mL Micro Spin Columns 50 100 - - Micro-Elute RNA Spin Columns - - - 50 96-Well Plate - - 2 - Adhesive Tape - - 4 - Collection Tubes 50 100 - 50 96-Well Collection Plate - - 2 - Elution Tubes (1.7 mL) 50 100 - 50 96-Well Elution Plate - - 2 - Product Insert 1 1 1 1 Documentation PROTOCOLS PI23600-15 RNA Cleanup Insert.pdf PI43200-1 RNA Cleanup Insert.pdf PI25100-8 RNA Cleanup 96-Well Kit Insert.pdf PI61000-3 _RNA_Clean_Up_and_Concentration_Micro_Elute_Kit.pdf SUPPLEMENTARY PROTOCOLS Supplementary Protocol - NGS Library Clean-Up and Concentration.pdf APPLICATION NOTES rna_cleanup_and_concentration_kit_effect_of_microrna_gc_content_on_rna_purification_efficiency_application_notes_66.pdf App-Note-100-Effects-of-microRNA-GC-Content-1.0.pdf SAFETY DATA SHEETS 23600_43200_SDS.pdf 25100_SDS.pdf 61000_SDS.pdf POSTERS Poster-16-Effects-of-The-Adenovirus-Type-5-Tripartite-Leader-Sequence.pdf Poster-27-High-Expression-Level-of-Hepatitis-C-Virus-E1-and-NS3-Genes-Under-The-Control-of-The-Adenoviral-Major-Late-Promoter.pdf Poster-26-Effects-of-Adenovirus-Superinfection-On-Transgene-Expression.pdf FLYERS RNA Clean-Up and Concentration Micro-Elute Kit.pdf 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 Powered by Bioz See more details on Bioz Related Services NGS RNA Isolation Related Products 100b RNA Ladder 1kb RNA Ladder CleanAll DNA/RNA Clean-Up and Concentration Micro Kit RNase-Free DNase I Kit Total RNA Purification Kit
RNA Clean-Up and Concentration Kits For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol For research use only and NOT intended for in vitro diagnostics. Powered by Bioz RNA Clean-Up and Concentration Kits For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol SKU 23600 Format Spin Column High Throughput Micro-Elute Size 50 Preps 100 Preps Contact Distributor Request sample