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Old 12-20-2012, 02:22 AM   #10
Junior Member
Location: Israel

Join Date: Dec 2012
Posts: 8

I did another run of sonications. I used Covaris shearing buffer (as suggested by Hamid) and tried 3 setups (each sample is 130ul, ~2.5*10^6 cells):
duty cycle 2%, 8min
duty cycle 5%, 8min
duty cycle 10%, 8min.

Cleaned the DNA and ran on gel. Got the same results for all 3 setups: prominant band in the 1000-3000 rangs. It seems the duty cycle doesn't change my shearing.
Any suggestions? Increase the time (with 2%duty cycle I didn't notice difference between 8 & 12min)? Increase the Peak incident power (currently at 105watts)?
I'm at loss. It shouldn't be this hard.
Gel is attached.
Thank you!
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File Type: jpg 20.12.12 covaris 8min 2-5-10%.jpg (41.9 KB, 33 views)
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