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-   -   Sonication innefective? (http://www.molecularstation.com/forum/chip-chromatin-immunoprecipitation-forum/70797-sonication-innefective.html)

Arastho 10-01-2009 11:06 AM

Sonication innefective?
 
Hello everyone,

I'm not sure what is happening with my procedure so I need your ideas if you please.

I'm working with 25x10^6 293T cells concentratred in 1mL. I need this amount to the ChIP assay.

When I go to sonicate, and I'm testing different conditions, I only get smears higher than 1000bp.
I tested different cells concentration: 12x10^6 and 5x10^6
I'm working in the maximum power p/10 seconds avoiding overheating the samples.
I didnīt test the % duty cicle, but I will...

But anyone have another idea? I need 200bp fragments.

Thanks

Arastho 10-15-2009 12:28 PM

Re: Sonication innefective?
 
Hello everyone,

For the 61 views until today and no answers, here is the answer:

Is the formaldehyde %.
I'm using 1%.

I tested 0,2%; 0,4%; 0,6%; 0,8%. With 0,8% i get my 200 bp bands and the CHIP worked.

Hope that this could help anyone.


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