Cutting M13 dsDNA

[nathan]

>>> On 4 Nov 1995 06:07:35 GMT, tedm at darkwing.uoregon.edu (Ted Michelini) said:


gareth_p at vcp.monash.edu.au (Jo Caine) wrote:

>> I have just completed a mutagenesis experiment using M13 vector and
>> I'm having trouble cutting the dsDNA of the M13 with the Nde/Bam.
>> I have isolated the dsDNA with a homemade resin based miniprep.
>> Does anyone have any suggestions of how to get cuttable dsDNA?  Do
>> I have to go to CsCl preps?
>> 

I haven't had this problem in any great fashion, sometimes I have
observed partial cutting.  I culture for 5-6 hours, then do a qiagen
plasmid spin column prep.  No extra steps. You can do a five mL
culture with no problems, as your cell density (at least with XL-1)
won't be that high.  Yes, the yield is low, but since I started molbio
working with M13, I am used to it.

nathan

--
Nathan O. Siemers, Ph.D.	
Bristol-Myers Squibb Pharmaceutical Research Institute
3005 First Avenue, Seattle, WA 98121
(206) 727-3741  	siemers at bms.com