Dear Hans Faix and Alan Kimmel,
                              I have used the pLPBLP plasmids for
generating single gene ablations. The method altogether is working
fine and I generated several gene ablations with this method. But I
faced some problems while PCR screening for the positive clones (for
gene ablation). I wanted to confirm Bsr cassette insertion from 5' &
3' orientation, hence I decided to design some common primers in the
Bsr region (Forward primer in case of PCR 3' orientation and reverse
primer for the 5' orientaion PCR). Out of several primers I designed
only one or two were working. If you (or somebody else) have some
bonafide primers in the Bsr region, could you please include this
information.

Thank you,
Ranjani

On 2/28/12, Alan Kimmel <[log in to unmask]> wrote:
> Colleagues,
>
> Hans has modified the protocols for our original cre-loxP mutagenesis
> system for better efficiency and we will be including this in
> Ludwig's & Francisco's new methods volume.
>
> We would be very happy to hear from everyone who has used the system
> successfully, with references noted, but also from those with
> queries, suggestions, etc. so we may best update it.
>
>
> with best wishes,
>
> Hans Faix   		[log in to unmask]
>
> and Alan Kimmel  	[log in to unmask]
>


-- 
Ranjani Dhakshinamoorthy,
Doctoral Student,
Christian-Albrechts University of Kiel,
Germany.