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.
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