This is a shot in the dark, but does anyone happen to love coffee and order lab reagents in bulk?

[Avery Ashley]

So with the help of you guys I'm just getting to the finishing touches of putting my lab together and it's about time to start actually using all this expensive equipment. 

As I'm learning, it can be quite expensive to keep a lab stocked with all the necessary supplies (enzymes, gels and the like), especially when you're quite small and can't order in bulk.

Anyway, I roast coffee as a small business, meaning that I get green coffee beans straight from farms all over the world and "cook" them so they're ready to brew with.

Now I'm just putting this out there for the heck of it, but if anyone would be interested in trading small quantities of lab reagents for freshly roasted coffee I'd be very excited to do so. 

My coffee retails for $12 a pound, but I'd be more than willing to trade a pound of coffee for like $6 (roughly what I buy it at) in supplies so that you come out on the top side. 

I'm not sure if this is viable for anyone on here, or if anyone would even be the slightest bit interested, but why not ask, right?

Of course, I take great pride in my coffee. I get it from the best farms in the world and put extensive time and effort into every pound I roast to ensure it's of the highest quality. I can roast as much or as little as you like, no quantity is too small and no quantity, within shipping constraints, too large. Every pound would be roasted to order the day of shipment as is my policy. 

In closing, I certainly don't want it to seem like I'm asking for a handout. I'm simply offering my services in return for yours. 

[Dakota Hamill]

Sounds like a neat idea, what are you immediately looking to do?  Did you manage to get everything necessary for PCR and running a gel with gel visualization?  If so I have some primers for fungal and plant ID that might interest you.

You can find enzymes and agarose and things in pretty small quantities, no need to buy 10,000 units or 500g really.  200 units for $50 of Taq and $30 for 25g of agarose will each last you ~ 50 PCR rxns and 50 mini gels.

[Daniel C.]

On Thu, Feb 21, 2013 at 8:39 PM, Avery Ashley
<avery...@rams.sccnc.edu> wrote:
> Anyway, I roast coffee as a small business, meaning that I get green coffee
> beans straight from farms all over the world and "cook" them so they're
> ready to brew with.

What's more interesting to me is to try to "process" the green coffee
beans with lactobacteria (and maybe a few others? I don't know what
we'd want to use) in an attempt to reproduce in a lab what happens in
the gut of civet cats naturally. You know, those things in southeast
Asia that eat ripe coffee cherries and then poop them out later.
Apparently the bacterial process in their gut changes the flavor
profile quite a bit. If someone could figure out how to do that
artificially I bet you could make a lot of money.

-Dan

[Avery Ashley]

As to what I'm immediately looking to do I don't really know. Right now I just need to learn how to use all these things I've bought haha. My overarching goal has been to preform a successful bacterial transformation before the end of summer, so I guess I need to work my way to that. 

As far as PCR goes I have everything but microtubes and a reason to do it. And as of next week I'll be able to run a gel, but I don't have a visualization system yet. 

Really, right now I'd be interested in anything just so I can get up and running and get my procedures down.

Where do you buy your stuff from? Those prices aren't bad.

[Avery Ashley]

That's a really cool idea. I'm going to look into that.

[Daniel C.]

On Thu, Feb 21, 2013 at 10:16 PM, Avery Ashley
<avery...@rams.sccnc.edu> wrote:
> That's a really cool idea. I'm going to look into that.

I think the most difficult part would be identifying the bacterial
profile of the civet cat's gut. I suppose you could just fly there,
shoot one, and empty its intestines into a beaker, but I'm not sure if
that's really the most efficient way to go about it...

-Dan

[Avery louie]

I would buy some gelgreen from phenix research- they have "sample" sizes for ~$20.  Gelgreen is stable at room temperature, and fluoresces under blue light.  I reccomend getting some cheepo LEDS from amazon- they have a lot of blue led strips for really cheap.  hook them up to like, a 1k resistor and a 9V and you should be in business.

As for a transformation- that is easy.  you can buy a kit from carolina and do it, or buy a bunch of pieces from all over and do it.

--A

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[Avery Ashley]

I briefly looked into it and it looks like its already been done by the university of Florida. It's a shame, that would have been cool work.

[Avery Ashley]

Cool thanks, I'll take a look at it.

[leaking pen]

There are already engineered enzyme soaks out there.

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[Nathan McCorkle]

I'd be interested, as long as you're a good roaster! I don't have
anything to trade right now, but I'll keep it in mind.

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[Eugen Leitl]

On Thu, Feb 21, 2013 at 10:10:05PM -0500, Daniel C. wrote:

> What's more interesting to me is to try to "process" the green coffee
> beans with lactobacteria (and maybe a few others? I don't know what
> we'd want to use) in an attempt to reproduce in a lab what happens in
> the gut of civet cats naturally. You know, those things in southeast
> Asia that eat ripe coffee cherries and then poop them out later.

The keyword in above sentece is 'ripe'.

[Koeng]

Here is some advice to get some free stuff :D 

go to www.mobio.com

They have a ton of samples... although I don't know if they will ship to residential address (I have made it a habit of shipping to business address) They have LB agar, broth, plasmid purification kits, gel purification kits and more! All for free! I used that when I was getting into transformations.

 Also I have something that might be helpful that actually came from indiebiotech :) check this out for your transformation-

https://github.com/cathalgarvey/biohacking-protocols/blob/master/E.coli%20Transformation%20With%20PEG%2BMgSO4.md

I would actually recommend this more then the kit because you don't run out of the CaCl mixture (I mean I bought a kg of it but it takes too long to make the concentration right) and all the materials come from your local stores! Then you can go on and use one of their mini prep protocols and purify your plasmid! When I started, I used this plasmid-  pGreen (No not the plant one)

I don't drink coffee often, but I would be willing to give all the help I can :)

Koeng

[Avery Ashley]

Thank you very much! That is extremely helpful information.

[Dakota Hamill]

Well if transformations is what you want to do, you should look into all of the necessary things to do that ie - plasmid of choice, selective growth media, competent cells, or materials required to make the competent cells, incubator, tools necessary for sterile technique.  I think a pGREEN or a pGLO would be a good start, moreso pGLO because it auto-fluoresces whereas pGREEN needs a UV light, but a UV light is good to have eventually for DNA visualization as well.  

You mentioned you got a good bit of lab equipment, what exactly did you get?

As for Taq, New England Biolabs is a few towns over for me and an amazing institution.  They were one of the first companies in the bio-tech area and make great products.

https://www.neb.com/products/e5000-taq-pcr-kit#tabselect0

That kit might be a good choice because it has your polymerase, buffers, dNTP's, and even comes with a ladder.  The kit allows you to modify PCR conditions as needed, by varying the concentration of your various buffers.  That's great if you have a handle on what you're doing, and could make for a really detailed learning experience by varying things to see their effects, but you might also consider a master mix.  

https://www.neb.com/products/m0270-taq-2x-master-mix#pd-properties-usage

The master mix has everything pre-mixed, so you don't have to do reaction optimization, but the downside is...you can't do reaction optimization because everything is already mixed together at a set concentration.  But for standard PCR, it should be OK.  I use the Taq 2X mastermix from NEB and it's worked fine for me so far.  Be aware of freeze-thaw cycles though.  I sometimes aliquot out enough master mix for 5-10 rxns in another PCR tube, and pull that one out instead of the master mix to avoid degradation.

I don't think you'll find Taq anywhere else for much cheaper, or if you do, it won't be THAT much cheaper, and might not be worth sacrificing quality to save $5-$10 when NEB produces some of the best stuff in the business.   On another note though, you might find it very difficult / impossible to get it sent to a residential address.

NEB also sells good ladders.

You can find Taq and other things like ladders and dNTP's from other sites, and they probably wont ask questions about where it is going to.

http://www.phenixresearch.com/

is where I've made my most recent purchases.

I've heard good things about MOBIO but havn't used them, they seemed to cut down on their samples a bunch, perhaps because it was being abused.

Epoch Lifesciences is also where I get all my spin kits, they're cheap and good.

Figure out exactly what it is you want to do and people can probably provide more direct advice.  I have pGREEN and pGLO I could give you as well as primers, but I have no bacteria for the transformation that would survive via shipping.

[Koeng]

I also wanted to add that the pGreen I use the bacteria literally turn green... so there is no need for UV light. I would also be happy to send you some :). My experience with pGLO isn't that great though... perhaps as a source of a promoter would be what I would use it for

another place for Taq would be openbiotech. I like them... they are "open source"

-Koeng

On Thursday, February 21, 2013 5:39:23 PM UTC-8, Avery Ashley wrote: