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Nuclei Prep Problems

histones chromatin yeast nucleosomes

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#1 jmiller623

jmiller623

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Posted 27 September 2012 - 09:52 AM

I prepared nuclei from S. cerevisiae and am trying to isolate chromatin in the form of oligonucleosomes. I sonicated the nuclei in a water bath sonicator for 1-12 cycles at high at 30s on and 30s off. When I treated the supernatent and pellet with Prot. K and RNase and ran them on a 1% agarose gel, I get a band around 700bp for both the sup. and pellet. There is no smear in the sup. gel but there is one for the pellet from 700bp up to above the 10000bp marker. The gel is shown in the post. The top is the sup. and bottom is the pellet.

Any ideas on what this band is? I ran a control that wasn't sonicated and got the same results for all of the cycles. I also ran the sup. and pellet on a SDS-Page gel and can see the histones after Coomassie.

Thanks!IMAG0495.jpg





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