How to optimize a nuclear extraction protocol?

19 September 2019 1 3K Report

Hi guys! I'm trying to extract nuclei from a lymphoma cell line, using a commercial kit. However, I suspect I'm lysing completely the cells during the first step (10 minutes on ice and then vortex 10 seconds), as I see something like DNA floating in the tube after vortexing. Also, after centrifugation (12000 rpm 1 minute 4°C) the pellet is flocked and not thick. How can I optimize this protocol? Maybe reducing the incubation on the ice at 5 minutes, without vortexing? or do I have to change the buffer dilution 1:9 or less and not 1:10 as indicated?

Muhammad Idrees

Catalog #: K266 Through this kit you can easily separate Nuclear/Cytosol proteins from cultured mammalian cells, as well as from tissues.

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Science method

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