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Old 03-07-2019, 06:21 PM   #15
ecbolo
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Location: California

Join Date: Mar 2019
Posts: 2
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Hi All,

I'm excavating this threat because my question is somewhat related to the above.

I am using a Covaris E220 to shear highly degraded DNA (from animal hair and museum tissues). The DNA starting fragment size between 600-3,000bp and I want ending sizes to be 300bp. The official Covaris protocol to get 300bp suggests the following for DNA > 10,000bp:

Peak Power: 140 W
Duty Cycle: 10%
Cycles Per Burst:200
Treatment Time: 80s

But, I've found suggestions that the closer in size the original DNA is to the desired post-shearing size, the longer the treatment time should be: https://covaris.com/resources/faqs/#toggle-id-100. Has anyone run experiments on the treatment time needed for degraded DNA? Should I run longer than 80s? Shorter? How much of a concern is over-fragmenting?

Thank you!!!!
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