[DIYbio] Re: Coolest open Source vector ever - anyone experienced at gibson assembly?

In addition, their plasmids lack complex features, common MCSs, and their standard sucks *unless* you have the parts registry which is about 3500$. The plasmids have lots of common restriction enzyme sites in the backbone (XhoI for example) which makes making compatibility layers for new parts very difficult.  The construction of said vectors to make a complex part takes a very long time, about 3 sequential cloning reactions from basic parts. While their plasmids do have some features that are useful like the terminators around their MCS, they didn't include the M13 primers, they included their own primers which companies don't have, making sequencing very large amounts of DNA more difficult. In addition, while there are negative selection methods they could use to simplify plasmid construction, they don't usually use them.

Pretty much, other than when competing iGem, their plasmids are a pain to work with. A lot of the projects there actually couldn't get prizes because their constructs didn't go in the standard plasmid so well. It's difficult to actually modify the backbone to fit your special needs unless you use gibson or the like. Plus they wouldn't accept plasmids in other formats. 

Because of these limitations, no labs I know of actually use it for normal cloning. They *all* use traditional cloning or golden gate, but mostly gibson. There's definitely need for an open source plasmid that actually fits all specifications well enough to be used.

-Koeng

On Monday, November 24, 2014 12:47:24 AM UTC-8, Michael O. wrote:

Hi Andreas,

Maybe you should take a look at http://parts.igem.org/Plasmid_backbones - there's no need to make yet another open source vector...

best,

michael