The isolation of RNase inhibitor from placenta in 1979 was one of the many contributions that have allowed modern molecular biology to examine genes and proteins in a way never thought possible. Now several types of RNase inhibitors are available to maximize the results of your RNA purifications, transcription or translation experiments, PCR amplifications, and cDNA syntheses.
While most biologists would love to rid all traces of RNase from their labs, the sturdy enzyme actually plays an important role in your body’s defense system. Found in bodily fluids, such as tears, mucus, saliva, and perspiration, RNase immobilizes invading bacteria and other microorganisms. These microorganisms also have their own brand of RNases, which are the chief complaint of researchers who use proteins and nucleic acids produced by, and/or derived from, prokaryotes. Allowed to exist unchecked, RNases end up in cell cultures and experiments in which RNA integrity is crucial.
To deal with the RNase, lab workers follow a list of precautions, including the use of gloves, dedicating pipettors exclusively to RNA work, and designating an RNase-free zone in the lab that is also shielded from air vents. In addition, RNase-free reagents, such as water and buffers, are widely available for purchase. Because these and other precautions can only go so far, researchers routinely use RNase inhibitors to block and/or denature RNases that may be lurking in their precious RNA or cDNA samples or other reagents.
RNase inhibitors, purified from tissue or produced by cloned bacteria, differ in the temperatures at which they’re active, the mechanisms by which they do their job, and the types of RNases, including RNase A, B, C, T1 and S1 nuclease, that they’re able to immobilize. Most companies offer inhibitors that don’t interfere with other enzymes that are important to the experiment, such as Taq polymerase, reverse transcriptase, or RNA polymerase.
Companies have developed inhibitors that are increasingly convenient to use. These include inhibitor-soaked towels—much like baby wipes—that are simply used to wipe pipette tips, counters, glassware, and other equipment and surfaces. Inhibitors are also available in spray bottles used to quickly and easily shower lab benches with a good dose of RNase decontaminant. For a quick check before embarking on an experiment, several RNase detection kits can make the procedure practically effortless.
You don’t have to endure the frustration of RNA degrading during key experiments. By simply choosing one of the many RNase inhibitors, such as those below, you can make discoveries that could help advance genomics to even higher levels.
Prime RNase Inhibitor is a protein of non-human origin that binds non-covalently and inhibits the same types of ribonuclease as HPRI, including RNases A, B, and C. Prime RNase Inhibitor can be used in cDNA synthesis, in in vitro transcription using the SP6, T7, and HeLa cell extract systems, and in in vitro translations using rabbit reticulocyte lysates and wheat germ extracts. Additionally, Prime RNase Inhibitor prevents RNA degradation in cell extracts prepared under non-denaturing conditions, and it may be utilized in first strand synthesis prior to PCR as well as in the RT-PCR of RNA isolated from single cells.
