Preparation of Sample

	BioSpec Products, Inc. (800) 617-3363 or (918) 336-3363

Preparation of Sample

Use 0.1 mm beads for bacteria, 0.5 mm beads for yeast, fungi and tissue culture cells and 1.0 mm beads for plant and animal tissue. Up to 400 mg wet wt of biomass can be disrupted per ml of liquid extraction media**. If you have solid tissue, pre-chop it into approximate 1mm cubes with a single-edge razor blade or fragment the tissue at liquid nitrogen or dry ice temperatures with a BioPulverizer™.

Fill either 2 ml screw-cap microtubes or 1 ml or 2 ml deep-well microplates one-half to two-thirds full with beads (BioSpec has a simple bead loader for 96-well microplates which makes loading easy and accurate, Cat. No. 762L, $96). Bead size and type should be matched to the biomaterial you are processing (click Here for details). Then add your extraction media and cells, being sure to fill the microtubes or wells almost to the top. Exclude as much air from the microtubes or wells as is practical. We recommend using screw-cap microtubes with o-ring seals. All snap-top microtubes release aerosol during shaking. Be sure there are no beads on the threads of the microtubes or on the lip area of the microplate(s) when sealing the microtube or microplate(s). If sealing microplates with a flexible mat or with adhesive film, firmly press the mat or film over the entire top surface of the microplate (BioSpec offers a high leverage mat press which makes sealing easy and secure, Cat. No. IT-EP-R, $1250).

Some microplates and their matching mats or sealing films work better than others. You may want to try different sealing techniques and microplates and mats made by several different manufacturers to find one that delivers the integrity needed to prevent sample leakage from well to well during the shaking process in the MBB-96. To date, we find that microplate products P-DW-20-C (2 ml) and P-DW-11-C (1.1 ml) with silicone rubber sealing mats AM-2ML-RD made by Axygen Scientific (Union City, CA) pass our dye test. As we receive feed-back from our users, additional acceptable manufacturer brands will be posted on our web site.

Operating the MiniBeadbeater

Insert microtubes into the tube holder, if applicable. Distribute the tubes symmetrically. Place the rack of microtubes or microplate (if using 1 ml microplates you can stack two at a time) into the chamber holder, top side toward you. Screw down the two black plastic knobs until the top stainless steel retainer plate is in contact with the microtube caps or top of the microplate(s). Tighten knobs snug but not over tight. Finally, screw down the white nylon wing nuts.

Turn on the “on-off” switch on the front of the MBB-96 cabinet and select the cell disruption time (0-5 min). Typical cell disruption times are 3 minutes.

During cell disruption, there will be a 10 degree raise in temperature per minute of operation. If you are isolating heat-sensitive material such as native proteins, consider pre-cooling your beads and sample, homogenizing for 1 minute only and then removing the vials or plate and re-cooling in ice-water for 1 minute. Cycle this way for a total “ON” time of three minutes. When extracting nucleic acids in nucleic acid extraction media, cooling is not needed.

Start homogenization by pushing the white button in the middle of the timer dial. The timer resets itself automatically to your selected disruption time at the end of the run.