DNA Shearing with S2/E210 Focused-ultrasonicator This Quick Guide provides DNA Shearing protocols when using microTUBE or miniTUBE and a Covaris S2 or E210 Focused-ultrasonicator. 130 μl sample volume- from 150 to 1,500 bp microTUBE AFA Fiber Snap-Cap microTUBE AFA Fiber Crimp-Cap 8 microTUBE Strip
50 μl sample volume - from 150 to 1,500 bp microTUBE AFA Fiber Snap-Cap microTUBE AFA Fiber Crimp-Cap 8 microTUBE Strip
200 μl sample - 2,000; 3,000 and 5,000 bp
Please note that miniTUBE requires removal of the Intensifier (PN 500141) from the E210 focused- ultrasonicator. Please see Appendix A for instructions. To fragment DNA to size 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. Values mentioned in this Quick Guide are nominal values. The tolerances are as f - Temperature +/-2°C Sample volume +/- 5μl - - Water Level  +/- 1
Sample preparation guidelines
- - DNA input: from 100 ng to 5 μg purified DNA 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. Recommended settings are subject to change without notice. See following link http://www.covarisinc.com/wp-content/uploads/pn_010158.pdf for updates to this document.
Supplies
The 500141 Intensifier is a small inverted stainless steel cone centered over the E Series transducer by four stainless wires.  The wires are held by in a black plastic ring pressed into the transducer well. If an AFA protocol requires “no intensifier”, please remove the Intensifier, using the following steps: 1. 2. 3. Empty the water bath.  Start the E System and start the SonoLAB software. Wait for the homing sequence to complete (the transducer will be lowered with the rack holder at it home position, allowing easy access to the Intensifier). Grasp opposite sides of plastic ring and gently pull the entire assembly out of the transducer well.  Do not pull on the steel cone or the wires.  The ring is a friction fit in the well – no hardware is used to hold it in place. The 500141 Intensifier (left) shown installed in the E System transducer well and (right) removed. Note the “UP” marking at t he center of the Intensifier. If a protocol requires the Intensifier to be present, simply reverse this process: Align the black plastic ring with the perimeter of the transducer well.  Note that the flat side of the center cone (marked UP) 1. should be facing up (away from the transducer). 2. Gently press each section of the ring into the well until the ring is seated uniformly in contact with the transducer, with approximately 2 mm of the ring evenly exposed above the transducer assembly.  Do not press on the cone or wires.  The rotation of the ring relative to the transducer assembly is not important. 3. Refill the tank.  Degas and chill the water before proceeding.
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