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Hi there,
I was trying to find some information to help me figure out what I could do to better shear some DNA. I found you advertising for qSonica while others are still using probe or Covaris.
I have been testing a few samples for which I would like to obtain fragment size between 300-350bp.
My setting were time varying from 30sec to 5min30sec, no pulse, amplitude 20% and temperature 4 degrees Celsius. Also my tubes are filled with 50ul of DNA (with low TE for a concentration ~287ng). When I run the samples after each addition of 30sec, it seems that I could get more 300-350bp for a total shearing time of 90s. However, I have a massive range (not so much of a pick) going from ~100bp to 700bp. If shearing for longer, the range is lower with more fragments lower than 300bp.
I was wondering if there were a reason for getting such results instead of a cleaner shearing (a real pick but of course there will be smaller fragment size).
Am I fouled by it and should I get much more when really increasing the time (e.g., 10mins?)
That will be fantastic if anyone had any tips as, unfortunately, I don't have much spare of DNA to test.
Thanks in advance.
I was trying to find some information to help me figure out what I could do to better shear some DNA. I found you advertising for qSonica while others are still using probe or Covaris.
I have been testing a few samples for which I would like to obtain fragment size between 300-350bp.
My setting were time varying from 30sec to 5min30sec, no pulse, amplitude 20% and temperature 4 degrees Celsius. Also my tubes are filled with 50ul of DNA (with low TE for a concentration ~287ng). When I run the samples after each addition of 30sec, it seems that I could get more 300-350bp for a total shearing time of 90s. However, I have a massive range (not so much of a pick) going from ~100bp to 700bp. If shearing for longer, the range is lower with more fragments lower than 300bp.
I was wondering if there were a reason for getting such results instead of a cleaner shearing (a real pick but of course there will be smaller fragment size).
Am I fouled by it and should I get much more when really increasing the time (e.g., 10mins?)
That will be fantastic if anyone had any tips as, unfortunately, I don't have much spare of DNA to test.
Thanks in advance.