RNA/Protein Purification Plus Kit

Overview

This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.

Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.  

This kit eliminates DNA efficiently using a gDNA removal column.

Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA.  The proteins will not require precipitation, resuspending of pellets, or any further cleaning.

This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.

Protocol

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Supporting Data

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Figure 1. Recovery of True Total RNA including microRNA from Hamster Liver. Panel A is a 1X MOPS 1% agarose gel showing the RNA that was isolated from 2 different samples of 10 mg hamster liver using Norgen's RNA/Protein Purification Plus Kit. Norgen's RNA/Protein Purification Plus Kit isolated both large RNA (represented by 28S and 18S rRNA) as well as small RNA with high integrity and without having to perform any additional protocol. Panel B is a result from a bioanalyzer resolution of the eluted RNA. Similar to the agarose gel, the Bioanalyzer showed that Norgen's RNA/Protein Purification Plus Kit has the added benefit of recovering small RNA. One microgram of the RNA was used in RT-qPCR reactions for the detection of human beta-Actin (for Large RNA) and miR-21 (for microRNA) genes. The RNA isolated using Norgen's RNA/Protein Purification Plus Kit showed superior recovery of both large RNA and small RNA including microRNAs as depicted by the successful miR-21 RT-qPCR (Panel C).

 

Figure 2. High Quality Total Proteins Eluted in Mass Spec-Compatible Buffer. Norgen's RNA/Protein Purification Plus Kit provides a column purification step for effective concentration and clean-up of the isolated proteins. The proteins are eluted into a buffer which is compatible with many downstream applications including mass spectrometry as well as standard protein quantification methods (including Bradford assays). In contrast, most competing multiple analyte isolation products require protein precipitation and the precipitated proteins are required to be resuspended in buffer with high-detergent content (such as SDS-PAGE loading dye) for full recovery. In this figure, the protein fraction isolated from hamster liver using Norgen's RNA/Protein Purification Plus Kit was resolved direclty on a 12% SDS-PAGE protein gel. Norgen's RNA/Protein Purification Plus Kit purified the proteins by column and the eluted proteins are already in buffer compatible with downstream applications.

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