( Trace amounts o...)Combat RNase Contamination in the Lab

Prevention
SUPERaseIn: The Ultimate RNase Inhibitor
SUPERaseIn RNase Inhibitor (patent pending) is a protein-based inhibitor of nonhuman origin that noncovalently binds and inhibits the most common and troublesome RNases including RNase A, B, C, 1 and T1. SUPERaseIn can be used in any application where RNase contamination could be problematic. It is ideal for in vitro transcription and translation, cDNA synthesis, RT-PCR, and preparation of RNase-free antibodies. Because it inhibits a broader range of RNases than traditional RNase inhibitors, SUPERaseIn is the most effective RNase inhibitor available, providing a higher level of protection against degradation (Figure 1).

Figure 1. SUPERaseIn vs. Placental Ribonuclease Inhibitor (RI). A ³²P-labeled RNA probe was incubated for 30 minutes at 37C in the presence of the indicated nucleases and either SUPERaseIn or RI. Both the SUPERaseIn and the RI were added at a concentration of 1 U/l.

The most widely used RNase inhibitor is placental ribonuclease inhibitor (RI), sold as Ribonuclease Inhibitor Protein. SUPERaseIn is distinct from RI in that it has a more robust interaction with RNases, does not require DTT for activity, and inhibits a broader range of RNases. While placental ribonuclease inhibitor is effective only against RNase A-type enzymes (e.g., RNase A, B, and C), SUPERaseIn also inhibits RNase I and T1. SUPERaseIn does not interfere with other enzymes such as RNA polymerases, reverse transcriptase or Taq polymerase. Additionally, SUPERaseIn is active up to 65C and at pH 5.58.5.

RNase-free Buffers and Reagents
It is essential that any buffer or reagent added to purified RNA be free of RNase contamination. A simple alternative to the tedious preparation of RNase-free solutions is to purchase certified RNase-free water, buffers, and reagents such as those offered by Ambion. After carefully isolating the RNA, it is crucial that the pellet be resuspended and stored in an RNase-free solution. Ambion's molecular biology reagents are tested under extremely stringent conditions for consistent lot-to-lot quality. Our quality control philosophy is to subject our reagent products to conditions 10-100 times more stringent than those experienced under normal use.

Detection
Sensitive Detection of RNases
If you need to detect RNases, then RNaseAlert is the kit of choice. This patent-pending technology reports RNase activity in a convenient and sensitive assay that delivers fast, accurate results in real time. The RNaseAlert Kit uses a novel RNA substrate tagged with a fluorescent reporter molecule (fluor) on one end and a quencher on the other. In the absence of RNases, the physical proximity of the quencher dampens fluorescence from the fluor to extremely low levels. When RNases are present, however, the RNA substrate is cleaved and the fluor and quencher are spatially separated in solution. This causes the fluor to emit a bright green signal (when excited by light of the appropriate wavelength) that can be readily detected by eye (with illumination on a UV box) or with a filter-based or monochromator-based fluorometer. The sequence of the RNaseAlert Substrate has been carefully optimized for sensitive detection of several RNases, including RNase A, RNase T1, RNase I, micrococcal nuclease, S1 nuclease, mung bean nuclease, and Benzonase.

RNaseAlert is provided in two convenient kit formats. The RNaseAlert Lab Test Kit is suitable for testing small sample numbers and can be used to ensure that solutions, tubes, tips, etc. are RNase-free. The RNaseAlert QC System is designed for high throughput assays in a 96 well format. This kit contains enough substrate to test 480 samples (5 x 96) using a fluorometer.

Elimination
Surface Decontamination
Common sources of RNase contamination include lab benches, pipettors, glassware, tubes, tips and electrophoresis apparatus. Ambion's RNaseZap provides an easy and effective way to decontaminate these surfaces. RNaseZap is not simply a detergent that spreads contamination around (like many other commercially available products), but contains three ingredients that are active against RNases. It has proved to be effective at removing contamination from glassware, plastic, countertops, and pipettors. RNaseZap is fast and convenient, destroying RNases immediately on contact. Simply apply the solution to the surface to be treated and rinse with nuclease-free water.

RNaseZap Wipes are a new and unique RNase control product designed for the elimination of surface RNases. RNaseZap Wipes are towelettes pre-soaked in an RNaseZap formulation, which allows RNases to be destroyed with a simple wipe.

back to top

Ordering Information
Cat# Product Name Size 1964 RNaseAlert Lab Test Kit 25 rxns 1966 RNaseAlert QC System 5 x 96 assays 2694 SUPERaseIn (20 U/l) 2500 U 2696 SUPERaseIn (20 U/l) 10,000 U 9780 RNaseZap 250 ml 9782 RNaseZap 6 x 250 ml 9784 RNaseZap 4 L 9786 RNaseZap Wipes 1 container (100 sheets) 9788 RNaseZap Wipes Refill 3 x 100 sheets
'"/>

Source: