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fresh and frozen liver tissue in NChIP using Medimachine


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#1 kblee

kblee

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Posted 01 November 2012 - 09:50 AM

Dear all,

Greetings.
I need to do lots of Native Chromatin Immunoprecipitation using both fresh or snap frozen liver tissue.
Since the clinical samples are precious, we will use Medimachine to homogenize the samples to achieve the nearly single cell suspension status.

Question time:
1. When using frozen tissue, suppose the cells dead. Can I merge the data using fresh tissue and frozen tissue?
2. Someone told me that there're tons of enzyme in the liver cells, when go through homogenizing procedure, the protein degrade extremely fast. So should I let the tissue go through the Medimachine procedure in frozen or let it thaw a little bit?
3. I know using tissue to perform ChIP is very difficult, not too much journal can be found in pubmed, why so difficult?
Anyone has experience on N-ChIP using tissue can share the pain and joy?

Thanks!




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