DNA shearing Sample Clauses

DNA shearing. The Covaris S-series single tube sample preparation system (Model S2) was set up. The Covaris tank was filled with fresh deionised water up to line 12 on the graduated fill line label, making sure that water covered the visible glass part of the tube. The chiller temperature was set in the water bath at 5 oC. The instrument was degassed for at least 30 minutes prior to use. A mix was prepared consisting of 3 μg of each DNA sample and 1X Low TE Buffer in a 1.5 ml LoBind tube to a total volume of 130 μl. A Covaris microTube was placed into the loading and unloading station and the cap was kept on the tube. With a tapered pipette tip 130 μl of DNA sample were transferred through the pre-split septa avoiding the introduction of any air bubbles into the bottom of the tube. The microTube is secured in the tube holder and the DNA is sheared according to protocol (Table 6). Setting Value Duty Cycle 10% Intensity 5 Cycles per Burst 200 Time 6 cycles of 60 seconds each Set Mode Frequency sweeping Temperature 4° to 7° C Table 6: Covaris DNA shearing settings Following the shearing process, the Covaris microTube was placed back into the loading and unloading station. With the snap-cap on, a pipette tip was inserted through the pre-split septa and the sheared DNA was removed and transferred into a new 1.5 ml LoBind tube. The process was repeated for each DNA sample.
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Related to DNA shearing

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