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RNAGEM™ PRODUCT OVERVIEW
The RNAGEM™ is the latest range of extraction kits developed by ZyGEM. The kits come in two configurations depending on the need to remove DNA (the RNAGEM PLUS kits come with RNAse-free DNAse I). Both kits produce RNA that is suitable for direct use in a wide range of downstream applications. The first kit to be released in the range is RNAGEM™ Tissue.
Most methods for extracting RNA rely on solid phase purification or solvent based extraction and precipitation methods. Often, lysis is achieved using agents or enzymes that must be neutralised or removed from the solution before the nucleic acid is usable for analysis. Alternatively they require modification of the downstream RT-qPCR reagents to allow for the carry-over. This requirement can increase the number of manipulations and limit your reagent options. In the case of solid phase or solvent-based methods, the manipulations may introduce bias into the population of mRNAs in the final sample. Essentially, every step that occurs between sample and assay introduces the opportunity to deviate form biological reality.
RNAGEM™ has been developed to offer a very rapid RNA extraction solution that minimises the level of manipulative bias that can adversely effect gene expression analysis. Unlike any other rapid prep kit, the RNA yielded by RNAGEM has been proteolytically treated, which significantly improves RT polymerase processivity yielding more sensitive and more reproducible results - especially with low abundance mRNA transcripts.
RNAGEM™ HAS MANY ADVANTAGES
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Closed–tube (protects sample integrity, minimises cross contamination and improves bio-safety)
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Hands–off (saves labor costs and allows for high throughput batch processing)
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One-step (reduces the opportunity for processing to alter the biological composition of the sample)
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Easily automated (compatible with most low cost, off-the-shelf liquid handling robots)
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Optimal for small sample sizes and low cell numbers (with linear yields from 10 – 100,000
cells. Using fewer cells provides robust and accurate data and allows the same culture to be tested for RNA expression and protein expression by western blot)
RNAGEM™ QUALITY CONTROL
RNAGEM enzymes buffers and reagents are rigorously quality controlled to guarantee no detectable contaminants. The QC tests are as follows:
- RNAse – none detectable (<10-10 Kunitz units per µl)
- RNA, DNA - Human, Bacterial and mtDNA – none detectable (<one genome)
- DNAse – none detectable (<10-5 Kunitz units per µl)
- Efficacy, activity and reliability
RNAGEM™ and RNAse contamination
RNAGEM is a powerful, broad-specificity, thermophilic proteinase that aggressively destroys ribonucleases, and no residual RNAse activity can be detected in extracts generated from human cell lines. The graphs below demonstrate RNAGEM™ powerful activity on RNAse A – a notably robust ribonuclease. A serial dilution of the RNAseA was made and one set of tubes treated left untreated (top) and the other set treated with RNAGEM™ for 5 minutes. As can be seen, all but the highest concentrations (which would never be encountered in biological samples) have been completely neutralised.
Rates of activity of serial dilutions of RNAse that are untreated (TOP) or have been treated with RNAGEM™ (BOTTOM). Y axis = relative fluorescence. X Axis = minutes, Z axis = number of Kunitz Units of RNAse A.
Click here for more information about RNAGEM Tissue
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