Total RNA Purification Kit

Product Description

Please note: This kit can be used instead of discontinued #53000-NB

• Isolate total RNA, from large rRNA down to microRNA (miRNA)
• Isolate high quality total RNA with complete size range from a variety of sources, even from plasma and serum
• RNA can be isolated and detected from as little as a single animal cell (see application note)
• No phenol or chloroform extractions
• Rapid spin column format, protocol completed in <20min
• CE-IVD marked version available
• Kits for large and small input amounts and 96-well kit for high throughput applications also available

This kit is suitable for the isolation of total RNA from a range of samples including cells, bacteria, yeast, virus and bodily fluids including plasma/serum, blood, saliva, CSF and more. Extract high quality and purity RNA with excellent RIN values and A₂₆₀/A₂₈₀ suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS and more.

The kit purifies all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol.  Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content.

Average Yield

For large serum and plasma input amounts (1 ml - 5ml) please use the Plasma/Serum Circulating RNA Purification Kit.

Performance Data

Fig.1. High Quality of Isolated RNA with Complete Size Range. Unlike most competitor's kits, Norgen's Total RNA Purification Kit allows for the isolation of all sizes of RNA, from the very large RNA down to the microRNA. Total E. coli RNA was analyzed on an agarose gel (panel A) and the Agilent® 2100 BioAnalyzer RNA Nano 6000 chip (panel B), respectively. Note the presence of small RNA species (red square) in the samples isolated via Norgen's kit and the absence of these RNA sp. in the comptitor RNA preparation.

Fig.2. Amplification of Both Large and Small RNAs in the same Extraction. Norgen’s Total RNA Purification Kit isolates the complete size range of RNA, including small RNAs without the use of phenol. Total RNA was isolated from 0.75 million HeLa cells using Norgen’s Total RNA Purification Kit, various silica-based competitors and phenol-based TRI Reagent. RT-PCR 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 used in a 20 µL qPCR reaction with primers against the human microRNAs (miR-19 and miR-21) and large mRNA (S15). Only the use of Norgen's Total RNA Purification Kit resulted in detection of both micrRNAs and mRNA amplification products similar to TRI Reagent but without the use of phenol.

Fig.3. Norgen’s Total RNA Purification Kit allows RNA isolation from a wide range of species or tissue types. Total RNA was isolated from 5 x 108 E. coli cells, 1 x 106 HeLa cells, 100 µL rat blood and 10 mg hamster kidney using Norgen’s Total RNA Purification Kit. One microliter of the 50 µL isolated RNA was analyzed on the Agilent® 2100 BioAnalyzer using an RNA Nano 6000 chip. Note the integrity of RNA from all inputs with the presence of small RNA species. Norgen’s Total RNA Purification Kit consistently isolates high quality RNA from various inputs that score a RIN value between 8 and 10.

Fig.4. Effective and Consistent Detection of miRNA from Plasma: Norgen's Total RNA Purification Kit can effectively isolate miRNA from plasma. Total RNA was isolated from 50, 100 or 200 µL of rat plasma in triplicates using Norgen's Total RNA Purification Kit (blue), a competitor’s silica-based kit (green) and a phenol-based RNA extraction method (red). Stem loop RT-qPCR using primers specific to miR-21 was performed. In brief, two microliters of the 50 µL isolated RNA was then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer or oligo dT primer. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21. Norgen's Total RNA Purification Kit is the only product that showed (1) consistent detection of miR-21 transcripts across all input volumes, and (2) Ct values correlated to input volume (decrease Ct with increase input).

Fig.5. Recovering Diverse miRNA Species from Plasma with Better Consistency. Norgen’s Total RNA Purification Kit isolates plasma RNA that performs more consistently in downstream applications such as microarrays. Total RNA including miRNA was isolated from 100 µL of mouse plasma in duplicate using Norgen's Total RNA Purification Kit, Competitor A's leading miRNA Kit or a phenol-based RNA extraction method. One hundred nanograms of extracted RNA from each kit was applied to an Illumina microRNA expression profiling kit. Scatter plots display better consistency (better clustering) of replicate signals from Norgen's samples. Genes with Pval < 0.01 for both replicates were in blue. The associated table suggested that Norgen's protocol recovered the same diversity of miRNA with better consistency (higher r2 value).

Fig.6. Better Diversity of miRNA Detected from Plasma. Norgen’s Total RNA Purification Kit isolates miRNA from plasma with better diversity than a leading competitor. Total RNA including miRNA was isolated from 100 µL of plasma using Norgen's Total RNA Purification Kit or 625 µL of plasma using Competitor A's leading miRNA Kit, and was applied to an NCode expression profiling kit. Microarray images suggested that Norgen’s Total RNA Purification Kit (left) isolates a better diversity of miRNA from smaller input amount of plasma than the competitor’s miRNA kit (right). Image courtesy of LC Sciences, Houston.

Fig.7. Norgen’s Total RNA Purification Kit allows sensitive RNA extraction from as little as a single cell. Total RNA was extracted from a decreasing number of 293 HEK cells from 1 million cells down to a single cell. Five microliters of the 50 uL isolated RNA was then subjected to a 20 uL reverse transcription using oligo dT primer. Three microliter of the reverse transcription was used in a 20 uL PCR reaction with primers to detect the human beta-actin transcripts. PCR products of beta-actin were detected from as little as a single cell. M is the marker lane, while the numbers correspond to the number of 293 HEK cells that were used for the RNA isolation procedure. Please view respective Application Note.

Related Products

Serum/Plasma RNA Purification Mini Kit (Spin Columns)