Any good way to remove DNase?
Posted 11 March 2009 - 08:09 PM
Posted 12 March 2009 - 03:08 AM
what kind of extraction method are you using?
In general to overcome this problem is to work fast during the extract protocol. keep the sample on ice. Use cooled reagents. Increasing the molarity of EDTA used would also help. These steps would slow down the plasmid degradation.
I would use phenol chloroform extraction to inactivate the DNAse.
Posted 12 March 2009 - 04:49 AM
Posted 12 March 2009 - 06:12 AM
Posted 12 March 2009 - 07:36 AM
ya i do miniprep as usual. i will try you method tommorrow. It seems that it is much more easier. Thanks a lot
Posted 12 March 2009 - 07:52 AM
my plasmid contain R6K replicon which only replicate in e.coli lambda pir strains (e.g S17 lambda pir and CC118 lambda pir), unfortunately, both strains are endA+. I really wish I had a lambda pir endA- host