freeze-thaw lysis of cells - do I have to add benzonase? (May/06/2006 )

i want to try this protocol
freeze-thaw lysis of cells
but it says add benzonase to digest DNA....i think we dont have it here...maybe i can add DNase but i had very bad experience with it and dont like it....so im wondering can I omit this step all together?

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Hi cathy,
You not giving detailes of your problems and for which purpuse you needed cells lysis by fridge and thaw methodes.So please tell its detailes then other people able to give you proper advice.

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hi
So i would recommend the traditionnal PMSF and also a protease inhibitor cocktail
i found that link interesting :
http://www.merckbiosciences.co.uk/Products...asp?catno=70746

for your exp, i think that you should do a DNases test in order to define the best quantity you need for your protocols.
Good luck kathy.
fred

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i want to do lysis on PC-12 cells and then WB.....

fred33, thank you a lot for the link! ill search again here in the lab maybe they have it....but of course i should figure out that DNase trouble since its available and ready to use ...maybe i should try adding Mg....

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