Cell Disruption: Select the Ideal Lysing Matrix
Mechanical homogenization using bead beating technology, such as MP Bio’s lysing matrices, is an effective method for cell lysis. Lysing matrices differ by size, shape, material, and composition.
When selecting the optimal lysis beads for your project, consider the toughness of your sample type, the particle size you wish to obtain, and the stability of your target molecule. MP Bio offers a complete solution for agricultural laboratories, including a large selection of lysing matrices for plant tissues and environmental samples.
Disrupting the Plant Cell Wall & Homogenizing Environmental Samples
Below are lysing matrix materials commonly used for plant cell lysis.
- Garnet matrix (Lysing Matrix A) can be used to lyse virtually any plant sample type as well as microorganisms and animal tissues and cells. It provides a high shear, high impaction, making it particularly useful for dense and elastic cell walls, but may be too aggressive for RNA isolation. This is not appropriate for environmental samples.
- Ceramic spheres (Lysing Matrix D and M) come in a variety of sizes and are suitable for extracting DNA, RNA, and proteins. Smaller ceramic beads are more appropriate for softer plant tissues (leaf, stem, flowers), whereas larger beads may be needed for certain plant seeds and needles.
- Stainless-steel beads (Lysing Matrix SS) generate high impaction with low shear, making them appropriate for extracting DNA, RNA, and proteins from bacteria, fungal spores, seeds, roots, and needles.
Environmental samples such as soil, manure, and compost have a complex composition and are often evaluated for microbial diversity. To optimize cell lysis for environmental samples, a combination of materials is ideal. Lysis Matrix E (ceramic, silica, and glass beads) is designed to lyse all microorganisms present in environmental samples.
Addressing Common Problems
Ultra tough samples that are difficult to homogenize: Dry grinding or cryogenic dry grinding may be necessary, but note that dry grinding can create an abundance of localized heat and cryogenic methods involve extremely cold temperatures — both of which can damage plastic tubes.
- Use metal lysing tubes and homogenizers that ensure thorough disruption in seconds.
- Consider using MP Bio’s Stainless Steel Lysing Matrix tubes and FastPrep® System.
Low yield from plant or environmental samples due to few microorganisms in the environmental sample, degradation by proteases or sample contaminants, or aggregation due to heat generation during bead beating.
- Use fresh soil samples whenever possible to avoid microbial loss.
- Check the aggressiveness of your lysing matrices.
- Use metal lysing tubes to avoid plastic contaminants.
- Chill your samples before and throughout the sample preparation process to avoid overheating.
Low molecular weight of target molecules due to the lysing matrix being too aggressive causing harsh molecular shearing by physical impact of beads.
- Consider using beads that produce a low shear, medium impact, such as lysing matrix E.