I learned hat some some of the old posts have been lost due to a server crash at bio-forum.
Well, there were some earlier discussions how to increase the the yield of plasmid preparations for Clontech's pTRE tight vector. This vector has a low-copy origin for E. coli, and even with the best tricks (e.g. adding chloramphenicol some minutes before cell lysis) , the yield remains marginal.
I wanted to bring the attention to a high copy version of this vector (pTREtight2), that is distributed by the non-profit org. AddGene for shipping costs (available only for Research labs). The vector is compatible to pTREtight, it differs only in the E. coli origin.
A description can be found here
http://www.molgen.mp...lmet/pTretight/
and here is the vector
http://www.addgene.o...dentifier=19407
and an useful positive-control plasmid
HA-GDP1 can be detected with any HA and most GAPDH antibodies (but has a slightly different size than GPADH, so you have the loading control directly on the same blot)
http://www.addgene.o...n...&cmd=findpl
Best wishes,
Markus
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