Who knows and uses the Plasmid pGag-PolGpt? - Poor plasmid yield with Maxi-Prep.... (Aug/11/2005 )

Hi,

does anyone know the plasmid pGag-PolGpt (published at Journal of Virology, April 1998, p1120-1124). We uses it, and i always have very poor yield at maxi-Preparation using the quiagen kit. From 250 ml bacterial culture I get only 250µg plasmid. Does anyone have an idea to improve my yield?

thx

H

I don't know that plasmid. But I'm working with several other HIV-plasmids myself and all of them are high copy number, so I assume this one is too?

What bacteria do you grow it in? Some give lower yields than others.

How long did you grow it, at what temperature, what speed did you shake it, did your medium look like it was fully grown (don' t know how to express this correctly)?

Use a richer medium than LB? There's for instance TB if I'm not mistaken, TB means terrific broth (just search for it on google).