low plasmid yields in dh5alpha

Ed Siefker via methods%40net.bio.net (by ebs15242 from creighton.edu)
Tue Feb 26 11:31:26 EST 2008


Thanks for all the help everyone.  I had our undergraduate run
10ul on a gel.  She got nothing but a faint smear in the image.
Too bad about that, but that's why we're doing this.
-Ed

Sudheendra Rao N R wrote:
> I guess you must be freshly transforming the DH5alpha with your old stock of
> plamids.
> Have you checked that plasmid on the gel? is it nicked? what about its
> concentration?
> 
> Sudheendra.
> 
> On Thu, Feb 21, 2008 at 6:48 AM, DK <dk from no.email.thankstospam.net> wrote:
> 
>> In article <mailman.723.1203535541.2451.methods from net.bio.net>, Ed Siefker <
>> ebs15242 from creighton.edu> wrote:
>>> Hi.  We've been going through our old plasmids and making
>>> fresh stocks.  We've found a few that we're not able to
>>> miniprep and get reasonable concentrations (<50ng/ul).
>> Are you sure these are not low/moderate copy number
>> plasmids to begin with? WIth plasmids bearing pBR322
>> origin, a "normal" miniprep almost never gives >50 ng/ul.
>>
>> DK
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> 
> 
> 




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