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Old 02-27-2014, 07:30 AM   #1
Location: Spain

Join Date: Feb 2012
Posts: 13
Default fragmentation of ovine DNA


this is the first time I will construct a DNA library to next-generation sequencing. I'm working with ovine DNA and trying to set up the fragmentation step in a Bioruptor Pico machine. I need a library with insert size of 350pb. I have tested several different conditions and 30"ON/30"OFF and 16 cycles is the best one. However, I have observed differences between replicates of the same sample, and also between different samples. In addition, I would need to get a closer curve in the Bioanalyzer in order to loose the least amount of DNA after the cleaning step. I have tried increasing the number of cycles to 18, but it doesn't work.

Can anyone suggest any idea? have you also observed this lack of reproducibility?

Laruo is offline   Reply With Quote
Old 02-27-2014, 07:59 AM   #2
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Location: Boston,MA

Join Date: Nov 2008
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The lack of reproducibility has more to do with the BioRuptor itself than with the DNA type.

We switched to a Covaris because of this very issue- it was not reproducible and very difficult to get exactly what you want.
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