This Quick Guide provides DNA Shearing protocols when using microTUBE-130, microTUBE-50, microTUBE-15, microTUBE-500, or miniTUBE and a Covaris S220 Focused-ultrasonicator.
Revision History

Values mentioned in this Quick Guide are nominal values. The tolerances are as follows: Temperature +/-2°C - Sample volume - microTUBE-15: from 15 to 20 µl, +/- 1 µl o microTUBE-50: 55 μl, +/- 2.5 μl o microTUBE Plate, Strip, Snap and Crimp Cap: 130 μl, +/- 5 μl or 50 μl, +/- 5 μl o microTUBE-500: 500 μl, +/- 10 μl or 320 μl, +/- 10 μl o miniTUBE: 200 μl, +/- 10 μl o Water Level +/- 1 -
Sample guidelines
- DNA input: up to 5 μg purified DNA (1 μg for the microTUBE-15; minimum 320 ng for the microTUBE-500) Buffer: Tris-EDTA, pH 8.0 - DNA quality: Genomic DNA (> 10 kb). For lower quality DNA, Covaris recommends setting up a time dose - response experiment for determining appropriate treatment times. DO NOT use the microTUBE or miniTUBE for storage. Samples should be transferred after processing. -
Instrument setup
Refer to the instrument manual for complete setup. - - microTUBE and miniTUBE have specific holders or racks associated with them.
Instrument settings
Recommended settings are subject to change without notice. - - Mean DNA fragment size distributions are based on electropherograms generated from the Agilent Bioanalyzer with DNA 12000 Kit (cat# 5067-1509), with the exception of the 320 μl microTUBE-500 protocol (High Sensitivity DNA Kit, cat# 5067-4626). DNA fragment representation will vary with analytical systems, please carry out a time course based on settings provided in this document to reach desired fragment size distribution. See http://www.covarisinc.com/wp-content/uploads/pn_010368.pdf for updates to this document. Part Number: Date:

55 µl sample volume - from 150 to 550 bp
microTUBE-50 AFA Fiber Screw-Cap (PN 520166)
To ensure reproducible DNA shearing, it is required to centrifuge samples before processing DNA in a microTUBE-15.  Please see Appendix A for instructions. 200 μl sample - 2,000; 3,000 and 5,000 bp
S220 Part Number: Date:
To fragment DNA to sizes larger than 5 kb, Covaris offers the g-TUBE: a single-use device that shears genomic DNA into selected fragments sizes ranging from 6 kb to 20 kb. The only equipment needed is a compatible bench-top centrifuge.
Additional Accessories

Technical Assistance
. . By telephone (+1 781 932 3959) during the hours of 9:00am to 5:00pm, Monday through Friday, United States Eastern Standard Time (EST) or Greenwich Mean Time (GMT) minus 05:00 hours By e-mail at techsupport@covarisinc.com
1. Sample loading and centrifugation microTUBE-15 AFA Beads Screw-Cap Load and centrifuge microTUBE-15 Screw-Cap as described before placing the tubes in the rack. If some of the sample splashes onto the wall of the microTUBE while removing from centrifuge or placing into rack, repeat centrifuge step. All liquid should be at the bottom of the microTUBE-15 before starting the AFA treatment. 2. Sample processing Use settings provided on page 3.
3. Sample recovery
Repeat the centrifuge step before recovering sample from microTUBE-15. Preparation Station and unscrew the cap Retrieve the sample with a narrow bore 20 μL pipet tip. It may be necessary to push the beads aside for full recovery