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Old 12-18-2012, 11:14 AM   #7
Hamid
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Location: San Diego, CA

Join Date: Sep 2009
Posts: 105
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Hi nfourier,

Most protocols and reagents used today for ChIP were developed and optimized for use with probe or bath sonicators. Since Covaris technology is quite different, different sample preparation reagents and protocol had to be developed. Based on your description, you are using a one step lysis/shearing protocol using the buffer you specified. Unfortunately one step/lysis/shearing protocols and reagents do not work with all cell lines using Covaris. The Covaris reagents and protocols are universal and will work with any mammalian cell lines and tissues. Our shearing buffer is quite simple. It contains 1mM EDTA, 10mM tris-HCl, 0.1% SDS. After shearing, you can easily add concentrated salts and detergents to the sheared chromatin to match the constituents of your IP buffer without the need to dilute.
I have attached a gel image of raw cells being processing using Covaris AFA.

Thank you

Hamid
Attached Files
File Type: pdf Raw mouse monocyte chromatin shearing.pdf (193.2 KB, 73 views)
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