Tough Tissue Homogenization with a Bead Mill
The quality of sample preparation often determines the quality of the results and the conclusions drawn from experiments.
Introduction
The quality of sample preparation often determines the quality of the results and the conclusions drawn from experiments. When it comes to homogenization, some specimens yield easily and others, such as extremely tough or very pliable plant and animal samples, can be very difficult to homogenize. There are many homogenizing methods available, including high shear, ultrasonic, blades and mortar-and-pestle systems. Bead mill homogenizers, like the Omni Bead Ruptor 24 (Figure 1), are very effective for processing multiple small volume samples.
The Bead Ruptor 24 from Omni International has a very broad performance range. It is gentle enough to deagregate cell material without breaking the cell wall, or powerful enough to process even the toughest samples, like pig aorta, rabbit cornea, peanut leaves and root material. Using sample tubes pre-filled with a variety of lysing beads, the Bead Ruptor 24 vigorously and uniformly shakes the tubes providing an efficient, consistent, high-yield and high quality homogenization. The unique motion of the beads assures maximum efficiency, thereby minimizing sample heating. Its powerful brushless motor assures the quietest processing possible. Processing beads are available in metal, ceramic, garnet, and glass. Up to 24 samples in 2mL tubes or 12 samples in 5mL tubes can be homogenized simultaneously with the BR24.
RNA Isolati on from Peanut Leaves
For years, researchers at the USDA’s National Peanut Research Lab had been searching for a fast and efficient method for homogenizing very tough peanut leaves. Prior to the introduction of the Omni Bead Ruptor 24, the most effective method they had been able to find was the use of purification columns. No previous mechanical homogenizers were capable of processing difficult peanut leaves. Frozen peanut leaf samples weighing between 0.1g and 0.15g were frozen prior to homogenization. Sample and buffer were added to each of the 24 2mL sample tubes that can be simultaneously processed with the Bead Ruptor 24. 2.8mm ceramic beads were selected for use with the peanut leaves.
The processing power of the BR24 was programmed to 5.5m/s for 2 cycles of 90 seconds with a 15 second pause between cycles. The advanced power and motion of the Bead Ruptor 24 led to significant increases in RNA yield. The previous method took 20 minutes to process one sample and led to a maximum collection of 20µg of RNA. The Bead Ruptor 24 led to collection of 20µg to 100µg of RNA in less than four minutes.
Conclusion
The advanced power and motion of the Bead Ruptor 24 from Omni International create a faster and more complete homogenization of tough sample material, such as peanut leaves. When asked about the Bead Ruptor 24, Phat Dang at the USDA National Peanut Research Lab said, “This machine is pretty amazing because it revolutionized the way we process plant tissue, especially difficult plant tissues. It has also allowed us to eliminate the need for costly commercial buffering compounds.” To learn more about the Bead Ruptor 24 and all of the other homogenizer solutions from Omni International, please visit our applications section at www.omni-inc.com/apps.