Mini-BeadBeater-Plus

 

IMPORTANT!  REMOVE AND DISCARD TWO PACKING SCREWS FROM THE BOTTOM BEFORE OPERATION.  SEE THE TWO ARROWS ON THE MACHINE.

 

POWER REQUIREMENTS.  The Mini-BeadBeater-Plus is a dual-voltage instrument (120v or 230v) and comes pre-set to 230v.  Determine the appropriate voltage at your location and set the switch at the back of the product.  Failure to select the correct voltage could result in irreparable damage.  Damage resulting from user error is not covered under warranty.

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The Mini-BeadBeater-Plus is a single-sample, high-energy bead mill designed to violently agitate microbial or tissue samples a sealed 2ml microvial containing tiny glass, ceramic (zirconia-silica or zirconia), or steel (chrome steel or stainless steel) beads.  The Mini-BeadBeater-Plus features a brushless motor operating at 4500 rpm and user-programmable time settings (0-99 seconds).  For complete product details see https:///products/mini-beadbeater-plus.

 

Typical Operating Conditions: 

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.  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 the sample and buffer.  The maximum amount of biomass that can be homogenized is 400 mg wet weight.  Finally, top up the microvial by filling it almost to the top with buffer or extraction media.  This last step is done to exclude as much entrapped air as possible when the vial cap is screwed on.  Note that the dry beads, especially the 0.1 mm diameter beads, can entrap air.  You may have to invert the microvial to wet the beads and then top up the vial with a little more buffer.

 

Firmly insert the vial into the arm assembly, the 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 seconds (the MBB+ automatically resets to the last time-setting used).  Depending on the amount and toughness of the sample, beadbeating 2 to 3 minutes typically disrupts over 90% of the tissue or cells.

 

Homogenate can be recovered by simply decanting after the beads have settled 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 in diameter or larger, inserting the micropipette tip through the settled beads to the bottom of the vial and aspirate the homogenate.

 

Bead Selection:  For a comprehensive bead selection guide and a complete list of lysis bead media visit  https://biospec.com/category/lysis-beads-guidelines.

 

Temperature Control:  After a 90 second run, the temperature of the cell homogenate will be approximately 10 °C warmer.  Cooling steps may be necessary when isolating proteins, membranes, or subcellular organelles.  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 not help cool the homogenate during the beadbeating process.

Temperature control is not necessary when isolating nucleic acids in aggressive extraction media containing phenol/chloroform, guanidinium salts, and/or detergents.

 Monitoring Lysis:  The MBB+ will disrupt over 90% of the cells after 2 to 5 minutes of operation.  Homogenization in a beadbeater involves a cell “cracking” action rather than other methods that use high shear.  When viewed under a microscope after beadbeating, empty but intact cell membranes may appear to the eye to be unlysed cells.  Therefore, to monitor cell breakage, rely on assay methods that measure released intracellular constituents (e.g., OD260, enzyme activity, protein staining, PAGE).