It'd be helpful to get the revised protocols ASAP rather than waiting for a methods book to come out. 

Best-

Ick

Richard A. Firtel
Professor
Associate Dean for Operations
Division of Biological Sciences
UCSD 
La Jolla, CA 92093-0380
Tel +1-858-534-2788
fax +1-858-822-5900

On Feb 29, 2012, at 6:32 AM, "Jan Faix" <[log in to unmask]> wrote:

> Dear Ranjani,
> 
> Thanks for your feedback. We can certainly list some Bsr primers in the
> chapter that work in our hands very well.
> 
> Best wishes,
> 
> Hans
> 
> *********************************
> Prof. Dr. Jan Faix
> Hannover Medical School
> Institute of Biophysical Chemistry, OE 4350
> Carl-Neuberg-Str. 1
> D-30625 Hannover
> Germany
> phone: +49-511-532-2928 (office)
> phone: +49-511-532-5922 (lab)
> fax: +49-511-532-5966
> e-mail: [log in to unmask]
> 
> *********************************
> 
> -----Original Message-----
> From: DICTY [mailto:[log in to unmask]] On Behalf Of
> ranjani dhakshinamoorthy
> Sent: Wednesday, February 29, 2012 11:25 AM
> To: [log in to unmask]
> Subject: Re: [DICTY - 759] cre-loxP
> 
> 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.