Re: [DIYbio] Where to buy restriction enzymes from ?

On Thu, 5 Mar 2020 at 13:12, Dakota Hamill <dkotes@gmail.com> wrote:

What I did years ago to start was open an LLC with my state government, then got my certification from the secretary of state and brought it to a bank, opened an account, and used that to make purchases.  It was out of pocket and it did suck to eat the $500 LLC fee in MA but we're one of the most expensive states, other states LLC's are much cheaper.    This is assuming you're in the states, but other countries I'm sure have cheap ways to make yourself a corporation.  Fake it till you make it! 

FWIW, Alaska's fee is $50, and you can use a registered agent for the in-state address, which usually runs another $100. This even works for people who live outside the US, and probably also works with other states like Nevada or Delaware. Besides the cheap registration fee, Alaska has no state corporate income tax, which simplifies taxes considerably. (Federal income tax still applies, though.)

Cheers,

—mlp 

On Thu, Mar 5, 2020 at 12:54 PM Tom De Medts <tdmedts@gmail.com> wrote:

Hi Cathal,

Thanks for your email.

However, I am not sure I understand what you are saying.

Restriction enzymes are proteins, so if one is trying to produce a certain RE from a plasmid that encodes it,

it would be necessary to have access to that bacterial strain, extract the plasmid from that strain and put it into your strain of intereste,

OR use that original strain itself to produce the enzyme, and then purify it.

All of this is logistically difficult in terms of strain procurement, 

and not the easiest protocols to successfully execute in terms of RE induction, purification and QC.

Which is why I prefer to buy REs from a supplier....hence my original email inquiring about RE sellers who would supply to DIY labs.

But if you meant something else, then I will look out for your clarification(s).

Thanks,

Tom

On Thu, Mar 5, 2020 at 12:41 PM Cathal Garvey <cathalgarvey@cathalgarvey.me> wrote:

Years back someone suggested using a restriction enzyme sample to transform bacteria and then streaking out on various antibiotics, to see if the production plasmid could be recovered as a contaminant.

Possibly it could make even more sense if you are willing to discard a miniprep-volume worth of RE mix and concentrate any contaminating plasmid first..

For best results though you'd probably want the higher end enzymes that have undergone some engineering

On 5 March 2020 17:38:01 GMT+00:00, Tom De Medts <tdmedts@gmail.com> wrote:

Hi folks,

For those of us in the US, working in a DIY / garage lab setting, rather than from an incubator or MakerHacker-Space, from where can we obtain restriction enzymes?

I looked at Amazon.com, and it looks like Takara Bio might have sold these at some point, but they are all unavailable at the moment.

Some tye REs of interest are:

• ECoR I, 
• BamH I, 
• Sma I, 
• Hind III, 
• Sal I, 
• Sac I, 
• Not I etc.
  

Thanks , in advance.

Cheers,

Tom

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