RNase: comparing thee to snow on a winter’s day

RNase: comparing thee to snow on a winter’s day

RNase contamination is a nightmare for any scientist working with RNA and controlling RNase contamination is not an easy ask, especially as the pattern of contamination is completely unpredictable, as the team at Phoseon Technology (OR, USA) discovered when they decided to put it to the test.

Using numerous sets of 96-well plates placed in various locations and left over the course of a day, the team observed that some wells would be completely clear of RNase contamination, and some would be completely contaminated. They concluded that it was completely sporadic, with no dependence.

“We expected it to be like the accumulation of snow on a winter’s day – steady and constant. But instead, it can be sudden, unexpected and catastrophic. It’s not easily defined. You could have no contamination for an hour of leaving equipment out, or it could be contaminated the moment you open a sterile package,” explained Theresa Thompson, an application scientist at Phoseon Technology.

It can therefore be easily concluded how RNase contamination would lead to increased waste, lost time and reproducibility issues. All three of these problems are incredibly timely issues when society, politics and sustainability are considered.

Working with Tris buffers

In any path of RNA research; if there is any degradation of the sample, results can be inconsistent, inaccurate and hard to come by. Though RNase is incredibly resistant to inactivation, usual routes of RNase decontamination include treatment with DEPC overnight and autoclaving, which is incredibly time-consuming.

Many experiments are carried out in Tris buffers. These buffers cannot be subjected to the same decontamination methods, as DEPC attacks the amino groups of the Tris, limiting its buffering capacity and destroying itself in the process so leaving it unable to inactivate any contaminating RNase.

Phoseon Technology discovered that a UV wavelength of 280 nm can target RNase A, leaving it irreversibly inactivated, with no drift on the pH or buffering capacity of the Tris buffers. Phoseon have since incorporated this into their own novel technology platform, KeyPro™, formed of high intensity LED-based lights that can reach intensities that haven’t been achieved with other light sources.

Immediately before running any experiment, the platform is switched on, and in less than 5 minutes, all equipment, including Tris buffers are completely decontaminated of RNase. The ways in which this platform can benefit researchers, can be drawn into a much bigger picture of the key issues for scientists at this current time.