Dear Fang,when I want to proceed with nuclei stored in Nuclei...

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Jennifer Deke
Aug 11, 2017
protocol Protocol: Nuclear Extraction from Arabidopsis thaliana
Dear Fang,

when I want to proceed with nuclei stored in Nuclei Storage Buffer, how do I get them out of it? Centrifuging did not seem to have worked well. Do you have a suggestion?

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Fang Xu Author Answered Aug 23, 2017

Shandong University

It's really odd, it never happens to me.you can try to spin longer time at1500g. But if you use higher speed it will break the nuclei. I don't know whether there's something differ for your reagent. You can try to add the washing buffer without triton to dilute it and then spin it. I never did it as I never have problems to get the nuclei. But I think this method can be tried.
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Jennifer Deke Answered Aug 23, 2017

Universität Hamburg

Hi Fang,

thank you for your answer.
Yes we did, and in the other buffers it worked, but not in the storage buffer. We resuspended the nuclei pellet in storage buffer, froze it in liquid nitrogen and stored it a few days in -80°C. After thawing it on ice we tried to centrifuge it, but we only obtained a rather smeary pellet after centrifugation at 20,000xg for 10 min. At less speed no pellet could be seen.

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Fang Xu Author Answered Aug 23, 2017

Shandong University

Hi Jennifer, thanks for your question. I usually centrifuge for 10min at 1500g to pellet the nuclei. And it works well for me, did you try it?.
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