IMPORTANT! REMOVE AND DISCARD TWO PACKING SCREWS FROM THE BOTTOM BEFORE OPERATION. SEE THE TWO ARROWS.
Note: The Mini-BeadBeater-Plus is a dual voltage instrument (120v or 230v) and comes pre-set to 230v. Select the appropriate voltage for your location using the switch at the back of the product. Failure to select the correct voltage could result in unrepairable damage. Damage resulting from user error is not covered under warranty.
The Mini-BeadBeater-Plus is a single sample, high energy bead mill designed to violently agitate microbial or tissue samples a sealed micro vial containing tiny glass, zirconia or steel beads. The Mini-BeadBeater-Plus features a single speed, brushless motor operating at 4500 rpm and two user programmable time settings (0-99 seconds). For complete product details see https:///products/mini-beadbeater-plus.
Typical Operating Conditions: Fill a 2.0 ml, screw-cap microvial at least 1/2 full (1/2-3/4 is okay) with beads. The rest of the microvial volume is filled with your sample and buffer. The maximum amount of biomass that can be homogenized is 400 mg wet weight. Tissue samples larger than a few 10’s of milligrams should be pre-chopped into pieces less than 1 mm in cross-section using a scalpel or single edge razor blade. Finally, the vial is filled to the top with buffer or extraction media. Try to exclude as much entrapped air as possible when the vial cap is screwed on. Note that dry beads, especially when using 0.1 mm diameter beads, can entrap air. You may have to invert the vial several times to wet the beads and then top-off the vial with a little more buffer.
Firmly insert the vial into the arm assembly, pointed end first. Close the blue safety cover. Select a time of 90 seconds on the MBB+ panel and start the MBB+. Repeat beadbeating for another 90 second (the MBB+ retains the last time setting used). This total beadbeating run time of 3 minutes disrupts over 90% of the tissue or cells. Shorter beadbeating times may be acceptable for sensitive analytical or PCR applications.
Homogenate can be recovered by simply decanting after the beads settle by gravity to the bottom of the vial. To maximize recovery of the entire product, one can either wash the beads with buffer or, when using beads 0.5 mm diameter or larger, plunge the micropipette tip through the settled beads to the bottom of the vial and aspirate the homogenate.
Bead Selection: For comprehensive bead selection guidance and a complete list of lysis bead media visit our web site: https://biospec.com/category/lysis-beads-guidelines.
Temperature Control: At the end of the 90 second run, the temperature of the cell homogenate will be approximately 15 °C warmer. Cooling steps are necessary when isolating proteins, membranes, or organelles. When isolating nucleic acids in aggressive extraction media containing phenol/chloroform, guanidinium salts, and/or detergents, temperature control is not necessary. To minimize heating, begin with a pre-chilled vial, beads, and buffer and operate the MBB+ for one minute. Then let the homogenate rest for one minute in an ice water bath. Cycle thus until homogenization is complete. Operating the MBB+ in a cold room does cool the homogenate during the beadbeating process.
Monitoring Lysis: The MBB+ will disrupt over 90% of the cells in 2 to 5 minutes of operation. The homogenization procedure involves cell “cracking” action rather than high shear. After homogenization, cell membranes may still appear to be intact when viewed under a microscope. Therefore, to monitor the time course of cell breakage, rely on assay methods that measure intracellular constituents (e.g., OD260, enzyme activity, protein staining, PAGE).