RNA/Protein Purification 96-Well Kit

(Cat# 37900)

For high throughput purification of total RNA (including microRNA) and total proteins from the same sample

Fast and easy processing using either a vacuum manifold or centrifugation
Sequentially isolate nucleic acids and proteins from a single lysate without phenol
Isolate total RNA, from large rRNA down to microRNA (miRNA)
High yields of isolated proteins
Ideal for small or difficult to obtain samples
Purify RNA/proteins from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants

Product Description
Product Description
Applications & Data
Applications & Data
Specifications
Specifications
Resources
Resources
Ordering
Ordering

Norgen’s RNA/Protein Purification 96-Well Kit provides a rapid method for the high throughput isolation and purification of total RNA and proteins simultaneously from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. Purification is performed using either a vacuum manifold or centrifugation. It is often necessary to isolate total RNA and proteins from a single sample, such as for studies of gene expression including gene silencing experiments, mRNA knockdowns or experiments correlating RNA and protein expression levels. Traditionally the RNA and proteins would be isolated from different aliquots of the same sample, however this novel technology will allow for their simultaneous isolation from the same sample. This will not only save time, but will also be of a great benefit when isolating RNA and proteins from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and proteins are derived from the same sample, therefore eliminating inconsistent results.

Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the process involves first lysing the cells or tissue of interest with the provided Lysis Solution. Alcohol is then added to the lysate, and the solution is loaded onto a 96-well plate. Under these conditions only the RNA will bind to Norgen’s resin, while the proteins are removed in the flowthrough (BIND 1). Next, an optional step can be carried out in which the genomic DNA can be digested allowing for a more pure RNA sample to be isolated. The bound RNA is then washed twice in order to remove any impurities (WASH 1). Lastly, the purified RNA is eluted into 75 µL of the provided Elution Buffer or water (ELUTE 1). Please see the procedure flowchart to the right. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). 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.

The proteins that are present from the initial flowthrough can now be loaded directly onto an SDS-PAGE gel for visual analysis. Alternatively, the protein samples can be further purified using the 96 well plates provided with the kit. After the RNA has been eluted from the plate, the flowthrough is then pH adjusted and loaded back onto the plate in order to bind the proteins that are present (BIND 2). The bound proteins are then washed with the provided wash buffer (WASH 2), and are then eluted such that they can be used in downstream applications (ELUTE 2). Please see the procedure flowchart to the right. The purified proteins can be used in a number of downstream applications including SDS-PAGE analysis, Western blots and mass spectrometry.

FEATURES AND BENEFITS

Reduce variablility - RNA and proteins are isolated from a single sample with no splitting of the lysate, thus reducing inconsistent results and variablility
Isolate from small samples - Simultaneous isolation of RNA and protein from a single sample. Ideal for precious, difficult to obtain or small samples such as biopsy material or single foci from cell cultures
Complete purification - The RNA and proteins are all column purified.
Fast and easy processing - 96-well plates can be processed in 25 minutes using either a vacuum manifold or centrifugation
Isolate a diversity of RNA species - All sizes of RNA are isolated, from large mRNA down to microRNA
Process a wide range of sample types - Isolate total RNA and proteins from cultured animal cells, tissue, blood, bacteria, yeast, fungi or plants

APPLICATIONS

RNA

Bioanalyzer
Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
RT-PCR for both large mRNA and small RNA including miRNA
Northern blotting
RNase protection
Primer extension
Expression array assays
Next Generation Sequencing
microRNA cloning
PCR-based virus detection
PCR-based viable bacteria detection

Protein

SDS-PAGE analysis
Western blots
Mass Spectrometry

Figure 1. Consistent Yield of High Quality RNA with Complete Size Range without the Use of Phenol. Norgen's RNA/Protein Purification 96-Well Kit allows for the consistent isolation of high quality RNA, with complete size range from the very large RNA down to small RNA without the use of phenol. HeLa RNA was isolated using Norgen’s RNA/Protein Purification 96-Well Kit in 15 replicates. The isolated RNA was resolved on a 1.2% formaldehyde agarose gel. All 15 replicates showed both high yield and high quality. In addition, all replicates showed effective recovery of all sizes of RNA including the small RNA (arrow).

Figure 2. Consistent Isolation and Purification of Total Protein from the Same Sample used for RNA Isolation. Norgen’s RNA/Protein Purification 96-Well Kit allows concurrent isolation of proteins and RNA from the same sample without splitting. Six of the flowthrough samples from the RNA isolation in Figure 1 were further purified using Norgen’s RNA/Protein Purification 96-Well Kit. Ten microliters of the 100 µL protein elutions were loaded on a 10% SDS-PAGE gel. The proteins are consistent in yield, intact and of the highest quality, and can be used in a number of different downstream applications.

Figure 3. Consistent Isolation of Both Large and Small RNA. Norgen’s RNA/Protein Purification 96-Well Kit allows for consistent isolation of both large and small RNA. Total RNA was isolated from samples of HeLa cells using Norgen’s RNA/Protein Purification 96-Well Kit. Aliquots of each total RNA sample were then used in 2 different RT-qPCR reaction. In the first RT-qPCR the S15 gene of mRNA was amplified (Panel A), and in the second RT-qPCR reaction the miR-21 microRNA was amplified (Panel B). Both the mRNA and microRNA were amplified in a consistent manner from all the samples, with low variability of the Ct values. Thus both types of RNA are being consistently isolated using this kit.

PROTOCOL

RNA/Protein Purification 96-Well Kit - Protocol
PDF version

PRODUCT INFORMATION SHEETS

RNA/Protein Purification 96-Well Kit - Product Information Sheet
PDF version

APPLICATION NOTES

POSTERS

MSDS

SUPPLEMENTARY PROTOCOLS

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