Hi, This is not typical. Try different cell lines for propagation and also add more sugar and glycerol to your medium - this helps.
Artem > Dear all, > > We are attempting to clone into pETDuet, pACYCDuet, pCola1Duet and > pCDFDuet > and we are encountering numerous difficulties. > > We are aware that most of them are low copy number, but even taking this > into account, we find it puzzling that whereas the first preps of the > vectors gave low, but significant amount of DNA, as time goes by the > quantities we get are constantly decreasing. And any subsequent step > (ligation, etc.) is a real challenge. > > Does anyone have any experience (succesful or unsuccessful) with these > vectors? > > Many thanks, > Silvia > > ------------------------------------------------------------------------------------------------ > Silvia Onesti > > Sincrotrone Trieste S.C.p.A. > SS14 - km 163,5 in AREA Science Park > 34149 Basovizza, Trieste ITALY > Tel: +39 040 3758451 > Mob: +39 366 6878001 > > Email: silvia.one...@elettra.trieste.it > http://www.elettra.trieste.it/PEOPLE/index.php?n=SilviaOnesti.HomePage > http://www.sissa.it/sbp/web_2008/C_Structural_Biology.html > ------------------------------------------------------------------------------------------------ >
