[DIYbio] Re: Openbiotech.com open source taq vector

The pdf they have of pOpenTag does contain the sequence of this plasmid, although in a very inconvenient format. I turned it into an annotated genbank file and put it up in case anybody wants it. (http://www.roningenetics.org/Sequencing.html) I also posted a slightly different, and to me, simpler, purification protocol for Taq I found a couple years ago based on the one they supply at the openbiotech website, it has fewer higher speed centrifugation steps (which I will use as the only way I have to get to 15000 rpm is in a microfuge), I have not tried either prep yet, I will try this one at some point. The comments in that protocol are those of the person who sent it to me, cant remember his name offhand.

Some of the descriptions of the plasmids on their website are not quite right. For pBR322 they say "pBR322 is a commonly used cloning vector. It contains a pUC origing, ampicillin resistance and supports blue-white screening." pBR322 is based on pMB1 (a ColE1 like plasmid), see Gene 2:95 for the original description of pBR322. It does not have a pUC origin and it does not offer blue/white selection. The need for a better way to select for inserts was a reason why the pUC plasmids with blue/white selection were developed in the first place. As to pUC19, a good reference is Gene 19: 259 (both of these older references are freely available) and its' origin and construction is more complex than having a "pUC" origin, it also involves sequence from some M13 phage vectors that had been developed for cloning and sequencing way back when. This is probably nitpicking, but those new to the field may get confused. If anybody from OpenBiotech is listening they might want to update the site a bit. They do deliver quickly. Hopefully Ill have some time in the future to make my own Taq now, this resource is very useful. If anybody at OpenBiotech wants some pBluesriptSSK+, another pUC like vector, I will be happy to send them some (and an annotated sequence file) if they want to offer that on their site. I think I have some pUC18 and either pUC8 or 9 stocked if they want either of those to offer also.
Tom