Re: [DIYbio] Re: The ODIN Needs Your Help - T4 DNA Ligase Testing - Free Ligase

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4453199/ It makes a lot of sense that these people recommend 19 base pairs. 

It's also definitely not just blunt end ligation, you can check out the data just in that paper, where they did control for that. I agree that I would need more controls to get decent results, I simply didn't think that it'd work this well. I'll update once I get those results (in the next few weeks I am definitely going to do more controlled experiments).

-Koeng

On Friday, October 30, 2015 at 1:22:57 PM UTC-7, Bryan Jones wrote:
I'm not an expert on reading patents, but it looks like the patent covers the method of the cloning procedure, not the production of SLiCE extract, nor the composition matter of SLiCE extract. Selling the SLiCE mix might not actually violate the patent (that'd be up to the user to do or not do). It also looks like the the patent only covers assembly of fragments that have "20 bp to 52 bp that are homologous". So using 19bp or 53bp homologous regions might be a way around the patent. Maybe a bit less efficient, but it looks like that would still work.
On Fri, Oct 30, 2015 at 3:03 PM Koeng <koen...@gmail.com> wrote:
Yea, my apologies for the non-controlled experiment, I didn't set it up to be very controlled because I didn't think I'd get that good of efficiency. I will be running a few more in the next few weeks so I will post the results. 

Unfortunately enough, it appears that SLiCE is patented ( http://www.google.com/patents/US20130045508 ), and the company that owns the patent charges an arm and a leg ( http://slicecloning.com/SLiCE.html , more than HiFi mix per reaction). I guess the ODIN can't sell the mix, but possibly sell a kit to make it yourself.

-Koeng


On Thursday, October 29, 2015 at 3:12:01 PM UTC-7, Josiah Zayner wrote:
This is pretty cool. If people try and repeat these experiments can they post detailed details(heh) of what they did so others can follow suite.

Also, it doesn't seem anyone is running the control of everything but the SLiCE mixture. If you are running an experiment can you do that and let us know how that goes?

Thanks,
     Josiah

On Thu, Oct 29, 2015 at 2:44 PM, Koeng <koen...@gmail.com> wrote:
I tried out my homemade SLiCE mix. I put 100ng of both insert and vector in for a 2 piece assembly, for 30 minutes. I compared it to some homemade Gibson mix.

On my plates, I got 13 colonies on the SLiCE plate and 42 colonies on the Gibson. I am extremely impressed with 1/3 considering just the simple strain and simple lysis procedure.

-Koeng

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