Integration of DNA fragments without restriction digest

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Fraser Moss
Fraser Moss's picture
Integration of DNA fragments without restriction digest

The link below is to a really useful protocol that I refined from a biotechniques paper by [url=http://www.ncbi.nlm.nih.gov/pubmed/11464525?ordinalpos=&itool=EntrezSyst...$=citationsensorl]Geiser et al [/url] to drop almost any fragment up to 2kb anywhere I like into any plasmid

http://www.scientistsolutions.com/science-protocols_displayprotocol_404d19b4-59b1-43ad-a40c-f2d99adf9bb9_e25959bd-687a-491c-a109-8b865e466c92.aspx

Jason King
Jason King's picture
This sounds like:
Fraser Moss
Fraser Moss's picture
The difference is that the

The difference is that the technique above works on circular plasmids. In-Fusion needs a linearized template into which you insert the fragment of your choice.

I've never pushed it to a larger fragment than 2kb so would not make a claim that it would work with a fragment >2kb without the experimental evidence to back it up, but you are welcome to try!

Jason King
Jason King's picture
I used to do more work with

I used to do more work with adenoviral plasmids and this would definately have been useful for these. I'll keep it in mind.