The Ice Nucleation Protein is part of an iGem distribution, so that part would not need to be synthesized. Most of the genes of interest could be cheaply cloned out of their species of origin, which could be had either through DSMZ or another culture collection or just asking authors of papers working with the bugs.
A basic bootstrapping set of enzymes might be the enzymes required for basic iGem standard assembly: thermostable pol, EcoRI, PstI, XbaI, SpeI, T4 ligase. Adding T4 DNA pol would be useful for SLIC cloning.
-- Re salvaging proprietary DNA from polymerase extracts, I had wondered if anyone had tried that one. How I would love to know the secrets behind Kapa's HiFi pol... I oughta try that trick out this week.
-Rob
On Tue, Nov 15, 2011 at 12:08 AM, Nathan McCorkle <nmz787@gmail.com> wrote:
That paper looks easy enough... the real investment is getting the
construct synthesized... just googling gene synthesis I found an offer
for $0.29/bp, so if you construct was 3000bp, that's $870
That's 100 people donating $10 each, not a terrible goal...
Someone mentioned due to inadequate enzyme cleanup, we might be able
to fish trade secret sequences out of commercial enzymes:
http://groups.google.com/group/diybio/browse_thread/thread/666d3b9f6cd439b2?fwc=1
(I couldn't find the older post I referenced in that thread, I'll try
searching some more)
What would a good set of enzymes for bootstrapping be? How much
synthesis do we need to buy before having a nice toolkit?
> --
On Mon, Nov 14, 2011 at 4:24 PM, Robert O'Callahan <ropoctl@rice.edu> wrote:
> The DIY bio community definitely needs cheaper tools and reagents. Would
> anyone have interest in having some strains suitable for the easy expression
> and purification of restriction enzymes, polymerases, and other useful ones?
> The method in this paper
> http://www.sciencedirect.com/science/article/pii/S1369703X11000325 seems
> pretty well suited for this. Another potential money saver would be a
> publicly available set of plasmids easily digested into quality DNA ladders.
> $50 per tube of restriction enzyme and $1 per lane of commercial ladders
> are not individually huge costs, but making these easy for individual to
> produce for themselves might lessen the barriers to entry.
> -Rob
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
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