SoniBeast
Performance optimization of the SoniBeast™ for your particular sample is highly recommended.
TIME STUDY: Conduct a beadbeating time study using control material. Try each of the six time settings on the SB (2, 5, 10, 20, 40, and 60 sec). For nucleic acid isolation, always beadbeat in an accepted nucleic acid isolation media. Good nucleic acid yields should be found in the range of 5-20 seconds when using 0.5 ml locking-top PCR tubes and 1-3 minutes when using 2 ml screwcap microvials.
BEAD AND VIAL SELECTION: Use 0.1 mm zirconia-silica beads for bacteria, 0.5 mm zirconia-silica beads for yeast, fungi and tissue culture cells and 1.0 mm or 2.5 mm zirconia-silica beads for fresh plant and animal tissue. In special cases such as grinding of dry leaf material, wet grinding of seeds, disrupting skin or cartilage, beads made of denser media such as zirconia or steel may be required. Additional guidelines on selecting bead media is available.
The Soni-Beast™ (SB) Beadbeater-type Cell Disrupter holds up to twelve 0.6 ml PCR tubes or three 2.0 ml microvials. Always use locking-cap PCR tubes or screw-cap microtubes with integral o-ring seals in order to eliminate possible aerosol formation or even ejection of the tube during beadbeating.
SAMPLE SIZE: Tissue sample sizes over a few tens of milligrams should be prechopped**. Up to 150 mg (wet wt) of biomaterial can be disrupted per 0.6 ml vial containing beads and extraction media. Up to 400 mg of biomaterial can be disrupted in 2 ml microvials.
** Note: Pre-chop solid tissue into pieces less than 1 mm in cross-section with a scalpel or single-edge razor blade. If your sample is large but has already been harvested and stored frozen, do not thaw. Rather, cryopulverize the sample. For information on cryopulverization see http:/products/40/biopulverizer/.
TEMPERATURE CONTROL: In general, start beadbeating with ice cold media and sample. During beadbeating the sample will be warmed. Warming will not effect the quality and yield of nucleic acids isolated in nucleic acid media, nor proteins isolated in urea- or SDS detergent-based media. However, if you are lysing cells to obtain native cellular components such as enzymes, antibodies, membranes, functional subcellular organelles or viable micro-organisms associated with tissue, feces or soil, close attention to heating during beadbeating is necessary. For each 10 seconds of beadbeating in the SB using 0.6 ml tubes, temperature will increase approximately 10 degrees. Heating in 2.0 ml tubes is slower (10 degrees per minute), but the cooler advantage is offset by longer times for complete cell disruption. Heating is best controlled by beadbeating for several short periods interspersed with a minute of manual cooling of the vial and its contents in ice/water mixture.
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PROTOCOL
1) Fill the tube or vial at least 1/2 full with beads. A approximate delivery of beads by eye is adequate. Then add up to 100 mg of cells and extraction media, being sure to fill the microtube almost to the top with media. For best grinding results, try to exclude as much air in the sealed vial as possible.
2) Load 1 - 12 locking-cap PCR tubes or 1 - 6 screw-cap microtubes into the SB vial holder table. Distribute them symmetrically as you would do with a centrifuge. Be sure to seat the tubes or vials all the way down into the rubber grommets.
3) Depress either the timer button (TIMED) or the manual button (PULSE). A typical time for complete cell disruption is 5-30 seconds using 0.6 ml PCR tubes or 2-3 minutes using 2 ml screw-cap microvials.