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

True true, EcoRI and HindIII are favorites of mine because of how much they supply. However, they are 6bp cutters... So I would actually argue that NotI and SbfI would be better for a new cloning system! I guess it depends on how much it would cost to remove the sites vs the actual enzyme cost.

I've noticed that at least at an academic lab people always use the same volume of enzyme... Like 1µl is 20 units of EcoRI but just 5 units of BtgZI... anyway it's a pretty wasteful approach, so if anyone is reading this is using restriction enzymes, go by units.

Then again, I haven't even cloned anything with classic restriction enzymes in a year or so because gibson is so good. However, gibson can cost an arm and a leg if you buy it directly from NEB (price for quality, their stuff gives me ~5-10xs better efficiency than our normal stuff). I know that SLiCE ( http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3333860/ http://www.ncbi.nlm.nih.gov/pubmed/24395368 ) would probably be very cheap to make in a DIY lab if created in mass, which might be useful for you. I'm curious what the efficiency is, but if it's good, I'd be willing to switch my home experiments over if it meant I wouldn't have to buy mixs anymore.

-Koeng

(btw: Thanks! I knew I forgot something)

On Thursday, October 15, 2015 at 10:27:12 PM UTC-7, Josiah Zayner wrote:

Yeah it is just a resale. Most people don't have ATP I imagine so I want to make sure it works as is, ya' know?

I will look at BsaI BsmbI but they are less used enzymes so they are freaking expensive.

You can do like 10 times the cloning with NcoI/EcoRI/HindIII on price maybe even more.

Same thing with Biobricks enzymes. I am selling those for iGEM folks but they are expensive. Someone should go back and start a cloning system that uses EcoRI and HindIII, the two least expensive enzymes.

Also, Koeng, you forgot to use the coupon code so I refunded you shipping!

On Thu, Oct 15, 2015 at 10:14 PM, Koeng <koen...@gmail.com> wrote:

How about the raw ligase? I buy ATP directly from NEB to do goldengates with, and some ligase from a non-commercial source might be interesting. 

Also, how are you getting the ligase? Expression of it in house? Or resale? Either or, I'm sure it'll be a great offer, but I am curious if you had the vector in house for some in vivo ligations I'd like to test out. Anyway, useful site! It's awesome that you've gotten more enzymes up, but I think BsaI/BsmBI are also a real important couple. Once you get materials to do GoldenGate, it's a pretty wonderful system, just wish I could get around to making some basic parts for DIY vectors...

-Koeng

On Monday, October 12, 2015 at 1:37:02 PM UTC-7, Josiah Zayner wrote:

My company, The ODIN(http://www.the-odin.com) is starting to ship restriction enzymes and ligase! However, enzymes are pretty heat stable. T4 DNA ligase is another matter, it requires ATP, ATP hydrolyzes. I need a few people to test out some T4 DNA ligase and reaction buffer that has been shipped. Shipping of Taq and other Master Mixes has worked and they contain dNTPs so I assume this should work out just fine but I need testers to make sure.

If you goto the website(http://www.the-odin.com/t4-dna-ligase-5ul/) and use coupon code FREETHELIGASE I will ship you some ligase for free as long as you promise to provide feedback on how it works after shipping.

Thanks,
     Josiah Zayner
     The ODIN
     ca...@the-odin.com

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