2 replies to this topic

[DavidA]

Hey guys,

I am looking for an effective way to disrupt mammalian white blood cells (physical shearing such as a beadmill is not possible at this stage) without actually purifying the DNA... As such i am considering a mild detergent such as 0.05-0.5% Tween 20 or Triton-X and perhaps some some mild vortexing.... Do you think this will be sufficient for this purpose? Another option might be simply several freeze/thaws however i don't know how efective this will be... Any ideas or suggestions?
Thanks

[knuf]

Hi,
For protein collection from HEK cells, I use a buffer with 0.5% NP-40 and do 3 freeze-thaw cycles with pretty good yield

[bob1]

Any combination of detergent and freeze/thaw will work - you won't purify the DNA unless you start adding salt and ethanol/isopropanol.  DNA will be released by any lysis step though, so sometimes the cells will get a little gloopy in texture - this can be broken up using DNAse or sonication.