[WHITE PAPER] Best practices for Cryogrinding with the Precellys Evolution

Cryogrinding, also known as freeze powdering, cryogenic grinding, freezer milling, and cryomilling, is the act of turning a deep-frozen sample into a powder in a temperature-controlled environment. Cryogenic grinding is one of the most effective methods to extract DNA, RNA, proteins, and metabolites from a biological sample.

In biology, most analytical workflows require a first step of homogenization to turn the sample into a liquid form compatible with the analysis technique. This homogenization step is usually followed by an extraction step in which the molecule of interest – such as DNA, RNA, proteins, or metabolites– will be isolated. Sample homogenization plays a crucial role in the yield and quality of the extracted molecules. There are several techniques for homogenization that can be divided into chemical, physical, enzymatic, and mechanical disruption methods. Mechanical lysis is now widely acknowledged as the gold standard for the homogenization of biological samples. For this reason, Bertin Technologies has chosen the 3D bead-beating technology to power its wide range of homogenizers, the Precellys tissue homogenizers.

An inadequate mechanical sample disruption protocol can lead to nucleic acid shearing or protein degradation. Proteins and RNA are thermosensitive molecules that can be degraded by the heat generated during mechanical homogenization. To address this challenge, Bertin Technologies has developed the Cryolys Evolution module, an automated cooling system that enables researchers to control the temperature of their samples during the whole homogenization process. To go even further, our team of scientists has designed stainless steel metal lysing tubes that allow users to snap-freeze their samples in liquid nitrogen prior to homogenization. In this white paper, we show how cryogrinding can be performed using the Precellys Evolution in combination with the Cryolys Evolution cooling module and the Precellys stainless steel metal tubes. We offer examples of protocols to efficiently pulverize deep-frozen animal and vegetable samples and maximize the recovery of nucleic acids, proteins, and metabolites.