Where to buy restriction enzymes from ?

[Tom De Medts]

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

[Cathal Garvey]

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

--
Sent from my Android device with K-9 Mail. Please excuse my brevity.

[Tom De Medts]

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

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[Dakota Hamill]

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! 

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[Christopher Monaco]

You might also reach out to local university labs or iGEM teams and see if they would be willing to share a little. Chances are they’ll have plenty of the common restriction enzymes. If you’re independent it’s great to develop a relationship with others who have the means to help you with harder to obtain materials and equipment.

[S James Parsons Jr]

Reach out to local laboratory managers from community colleges, universities and the like. Talk to them about your goals, and see what their recommendations are. Most will donate to your exploration.

Sent from my iPhone

> On Mar 6, 2020, at 7:34 AM, Christopher Monaco <cmona...@gmail.com> wrote:
>
> You might also reach out to local university labs or iGEM teams and see if they would be willing to share a little. Chances are they’ll have plenty of the common restriction enzymes. If you’re independent it’s great to develop a relationship with others who have the means to help you with harder to obtain materials and equipment.

>
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[Meredith L. Patterson]

On Thu, 5 Mar 2020 at 13:12, Dakota Hamill <dko...@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 

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[Tom De Medts]

Thank you all for your informative and helpful responses.
And sorry for my late response. I had a bad case of the flu ...

Dakota Hamill - thanks for suggesting registering entity as LLC to formally order lab items
Meredith L. Patterson - thanks for indicating that in some states it might be easier / cheaper to do this

Christopher Monaco - Thanks for advising me to connect with local univ. labs & iGEM groups
S James Parsons Jr - We have a local community college, I might talk to them about my goals

Looks like Promega supports select iGEM teams with reagents and other lab items. 

Anyone have 1st hand experience with this sort of engagement with a commercial supplier like Promega (or others like NEB, TaKaRa etc.)?
https://www.promega.com/resources/student-resource-center/igem/

Cheers,

Tom

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[Tom Randall]

Several years ago I got some enzymes from Promega delivered directly to my residence, but I haven't ordered from them in a while so I dont know if they still are selling directly to DIY people, as opposed to NEB, which will not sell directly.

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[everymanbio.com]

I'm in the process of buying restriction enzymes from NEB and recently had my account established as a sole-proprietor. All I had to do was create an account and wait for my account to be approved. It was internally routed to a sales rep that serves my area. She kindly reached out to let me know what I needed so she could get me a quote and get my order in.

Shipping is pricey unless the order is over $250. I looked at Takara Bio. While they'll support and ship to us just fine, I found their customer service to be rather disorganized. n=1 so maybe it was just an off day. Anyway, hope this helps.