low plasmid yields in dh5alpha

Jose de las Heras via methods%40net.bio.net (by josenet from tiscali.co.uk)
Thu Feb 21 03:36:45 EST 2008


"DK" <dk from no.email.thankstospam.net> wrote in message 
news:Xj4vj.2179$fU6.161 from newsfe02.lga...
> 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

Yes, I'm sure... they went back to the usual >300ng/ul once the lysis 
solution was taken care of.

They were all a series of constructs with the same backbone that we have 
worked with regularly, so a lysis problem was the first thing we thought 
off... and were right. If all troubleshooting worked like this! ;-)

Jose 




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