Hi all,
I am wondering if there's any paper or website with detailed information how to design a floxed gene, i.e. how to position your loxp sites, such as in the introns, avoid this and this sequence, be careful about promotors and enhancers if you want to flox the first exon, etc.
Another question related to this is, how does a floxed-Stop-codon sequence look like? Is it just loxp-TGA-loxp or is it more complex? Because one probably needs an ATG to start transcription and then to have a STOP codon right behind it.
Thanks for all your answers,
all the best.
Hello Andreas,
this presentation (http://www.betacell-cellbio.eu/teaching/LICENCE_ANGLAIS/Floxing%20In%20and%20Out.pdf) by Dr. Gawlik provides a nice introduction to floxing.
Best regards, Christian
Hi Christian, thanks for your reply. However, this is a nice introduction how it works with basic pitfalls. I am rather looking for a Methods paper how to actually do the floxing as mentioned in my question, i.e. proper engineering of the locus of interest.
I found this protocol: http://cdn.b-bridge.com/products/K005_Protocol.pdf
This paper might also be interesting for you: http://link.springer.com/protocol/10.1385%2F1-59259-178-7%3A175
I hope this helps you.
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