This Quick Guide provides DNA Shearing protocols when using microTUBE-130, microTUBE-50, microTUBE-15, microTUBE-500, or miniTUBE and a Covaris M220 Focused-ultrasonicator. 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 - microTUBE-50: 55 μl, +/- 2.5 μl - microTUBE Snap-Cap: 130 μl, +/- 5 μl - microTUBE-500: 500 μl, +/- 10 μl or 320 μl, +/- 10 μl - miniTUBE: 200 μl, +/- 10 μl •  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) •   Buffers: 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. •  WARNING: 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 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. 11
Table of Contents

130 μl Sample Volume: 150 to 1,500 bp
microTUBE AFA Fiber Snap-Cap Vessel
55 μl Sample Volume: 150 to 550 bp
Even if the water level check button is green in SonoLab, please check that water is in contact with itsert when using microTUBE-50. We recommend filling the water bath with 16 ml water. 44 15 μl Sample Volume: 150 to 550 bp microTUBE-15 AFA Beads Screw-Cap
To ensure reproducible DNA shearing, it is required to centrifuge samples before processing DNA in a microTUBE-15. Please see Appendix A for detailed instructions. Vessel 200 μl Sample Volume: 2,000, 3,000, and 5,000 bp
Vessel
microTUBE-500 AFA Fiber Screw-Cap Vessel
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. 7 Additional Accessories
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. 2. Sample Processing: -  Use settings provided on page 3. feel the magnets contact. lower the holder weight on top to keep the microTUBE in place. 3. Sample Recovery: Place microTUBE-15 in Preparation Station and unscrew the cap. Retrieve the sample with a narrow bore 20 μ pipet tip. It may be necessary to push the beads aside for full recovery. 9
Technical Assistance
•  By telephone (+1 781.932.3959) during the hours of 9:00 a.m. to 5:00 p.m., Monday through Friday, United States Eastern Standard Time (EST) or Greenwich Mean Time (GMT) minus 05:00 hours •   By e-mail at ApplicationSupport@covaris.com
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