Re: [DIYbio] Re: Experiment with DNA and learn the techniques and concepts in a convenient and easy way

       You literally need no equipment for the GFP cloning kit. If you have a lab you will use a water bath for the heat shock at 42°C and a 37°C dry incubator to grow the bacteria in the plates provided. Yet if you don't have these you can just mix hot and cold water until a thermometer tells you you've reached 42°C just before the heat shock in the transformation step, and as far as incubation goes there are two options we recommend, the easiest is tu place the plates behind your fridge where it gets warm and if you want to be more sophisticated you can just put a 40W old-school light bulb into a cardboard box the size of 4 six-packs and you'll reach a temperature close to 37°C (test with a termometer). 

Matías

On Thursday, March 8, 2012 9:29:41 PM UTC-3, Matías Gutiérrez wrote:

Sorry about the multiple e-mails, but I also forgot to mention that it includes a UV led lamp so that you can visualize fluorescence, it looks really bright, like this:

On Thursday, March 8, 2012 9:20:38 PM UTC-3, Matías Gutiérrez wrote:

Also included, forgot to add it into the list. :) but not for propagation, the propagation is achieved by plating the bacteria in the solid LB media. The liquid LB is used for recovery after heat shock transformation. 

Basically the idea is that you'll have everything you need, we've tested these in classrooms for over 3 years with great results and have improved our kit each year, this would be the first version that is not meant for the classroom but for enthusiasts! 

We could also provide individual components in a convenient ammount so that ppl don't need to buy bulk like research labs usually do. 

Matías

On Thursday, March 8, 2012 8:55:43 PM UTC-3, Nathan McCorkle wrote:

Matias, what about liquid LB for propagation?

On Thu, Mar 8, 2012 at 6:46 PM, Matías Gutiérrez
<matiasgutierrezm@gmail.com> wrote:
> Hey Guys,
>
>    First of all thanks to Edwin for your warm wishes :)
>
> Now regarding these kits, they basically include everything.
>
> For example the GFP cloning kit includes the following:
>
> plasmid encoding GFP and ampicilling resistance (GFP expression is induced
> by arabinose)
> bacterial stock culture (stab) that needs to be propagated before the
> experiment
> 2 plates containing propagation media (LB)
> plate containing selection media (LB + ampicillin)
> plate containing selection media and inducer (LB + ampicilling + arabinose)
> inoculations loops, microtubes, transfer pipettes, etc
> detailed protocol for the experiment
> Slide show with explanation of the protocol
> further reading material
>
> If we can work out a good distribuition network I'd really like to make this
> available for the community at a low cost (hopefully less than USD$30.00)
>
> Matías
>
> On Thursday, March 8, 2012 6:29:05 PM UTC-3, Avery wrote:
>>
>> You might just want to get pGREEN from carolina.  There is no need for
>> arabinose with that plasmid.
>>
>> --Avery
>>
>> On Thu, Mar 8, 2012 at 3:48 PM, Mega <masterstorm123@gmail.com> wrote:
>>>
>>> So you are still dependent on chemical companies   that may deny
>>> access.
>>>
>>> And if you (vacuum) destill the apple juice so the concentration gets
>>> higher? Is arab heat stable (say 100°C)?
>>>
>>> On 8 Mrz., 20:20, Derek <dere...@gmail.com> wrote:
>>> > Nope, no arabinose at the supermarket. It's found in low
>>> > concentrations in grape and apple juice, but I wasn't able to get
>>> > sufficient concentrations to enable GFP production.
>>> >
>>> > Best price I found, and nice people to deal with, are the folks at
>>> > Cascade Biochemicals,http://www.cascadebiochems.com/
>>> >
>>> > --Derek
>>> >
>>> > On Mar 8, 9:26 am, Mega <masterstorm...@gmail.com> wrote:
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> > > As for 2. GFP I want to mention, you need arabinose.
>>> >
>>> > > You can get it as DIY biologist, but not from the supermarket ( I
>>> > > think so, in fact never looked for arab there)
>>> >
>>> > > And you need a black light source.
>>> >
>>> > > There's a plasmid, pVIB, that is always 'on' and needs no special
>>> > > substrate. It may be more attracting to people if they see it glowing
>>> > > in the dark without UV-Lamps.
>>> > > That may only be true for me, at all. Though, I recommend thinking
>>> > > about this.
>>> >
>>> > > Best
>>>
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics