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sad and mad cloning

Leonard N. Bloksberg bloksber at pilot.msu.edu
Wed Jan 18 11:56:00 EST 1995

In Article <9501160240.AA94252 at lamar.ColoState.EDU> "xuhu at LAMAR.COLOSTATE.EDU (XuHu)" says:
> Dear Netreader:
>     We are having a great difficulty in cloning lambda genomic fragments(
> one 4.5 kb and one 14 kb) into pBluescript xho 1 site.  We grew the white
> transformants and checked inserts by restriction digest.  We found all of
> them (from both fragments cloning) are deleted pBS vector (about 2.2 kb)
> without inserts.  Check ligation there was shiftup on gel compared with
> self-ligated pBS. The gene we are cloning may involve in cell division of a
> plant.  Can anyone give some suggestion or comments.
>                                                      Xu Hu
I have had occational problems with the BlueScript vector series.  Sometimes
it seems to "mutate" into forms that do not clone or sequence properly, but
may still give apparently good digests on gels.  Storing your strains on
plates in the fridge is a good way to induce this problem.  Try a more stable
vector like pTZ or pUC.  I'm not knocking pBS, when it works it's great (and
I do use it), but if not handled properly it can be a hassle.
Good Luck
Lenny Bloksberg
bloksber at pilot.msu.edu

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