RNA Hygiene (Part I) – RNases and Your RNA Hygiene (Cleaning Solutions)

Posted:  04/15/2019

 

RNA Hygiene Microblog Series

Introduction

Novel techniques are being developed every day for use with RNA to help achieve more insightful results. This monthly microblog series will share an array of tips and techniques for hygienically working with RNA that the molecular lab newbie, as well as the 30-year RNA veteran researcher, will find helpful. Here at Advanced Biotechnologies we are cheering you on as you work to take extra care and precaution to ensure correct RNA experimentation outcomes. We hope you enjoy this series.

RNA Hygiene (Part I) – RNases and Your RNA Hygiene (Cleaning Solutions)

The primary enemies of RNA are the RNA-degrading enzymes known as RNases. RNases are found ubiquitously in the environment, including many of which that are found on human skin. For this reason, gloves should be worn at all times when handling RNA, associated labware, and equipment. Also, be sure to pull the glove up over lab coat cuffs as the wrist area is one of the most commonly overlooked sources of RNase contamination. To help keep your lab coat cuff in place, if your sleeves are long enough, try cutting a whole in the cuff for your thumb to fit through.

In the lab, the implementation of a clean dedicated area for RNA that is isolated from other lab activities is always a good practice.

Some RNases are so hearty that heat, even at boiling temperatures, is not sufficient to eliminate them. For this reason, chemical inactivation is used in place of normal heating, microwaving, or autoclaving. Many commercial solutions are available which effectively inactivate RNases (RNase AWAY, RNaseZap, etc.), but they are often prohibitively expensive. Fortunately, many lab-ready recipes are available on the web. Use these with caution, however, as most contain hazardous chemicals. Wipe down all surfaces and equipment first with the RNase inactivating agent, followed by RNase-Free water, and then finally 100% isopropanol. Be sure not to spray the RNase inactivating solution directly onto pipets, but rather use a clean paper towel to apply and wipe down. The follow-up applications of water and isopropanol are necessary to remove any residual RNase inactivation solution as it can cause RNA degradation itself, is hazardous, and is often corrosive to metal surfaces.

This RNase removal method is great for equipment, glassware, and counter surfaces, but what about inactivation of RNases in materials coming into direct contact with the RNA such as buffers and reaction mixes? Stay tuned for Part II of our series where we discuss the use (and misuse!) of RNase Inhibitor protein!

 

RNaseZap link: https://www.niehs.nih.gov/research/resources/assets/docs/instrument_cleaning_rnase_decontamination_508.pdf
RNase Away Link:
https://www.bio-world.com/site/accounts/masterfiles/MSDS/MS-10730002.pdf