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

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.

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.

Best

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?.