Rescue from DNAase activity - (Mar/15/2006 )

Hello everybody,

I had prepared a maxi prep of my plasmid and I think it has DNAase activity, once I start to restrict I see white streaks on my gel.

Is there a way that I can rescue my plasmid from DNAase activity...


Regards,

Harli

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A phenol/chloroform extraction followed by a chloroform extraction followed by an ethanol precipitation will remove most proteins, including DNAses.

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Also next time in your maxi prep, don't forget to treat the Buffer (PB in qiagen, buffer to inactivate DNase) twice. Sometime one treatment is not enough.

Also check the contamination with your buffers including elution buffer(or DW) and the buffer for restriction enzyme . In many cases, DNase is from the buffer contamination, but not from the cells.
If possible, change the buffers into new ones from next time.

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