Science

ALHAMDULILLAAH i am pass... in my MIT

Im sorry, I do have the plasmid map. It seems that he was referring to the attached document he had sent in the email but I never saw it. I have attached the plasmid map to my post. 

On Tuesday, July 31, 2012 11:19:26 PM UTC-4, Chowe wrote:

That is great to hear! I have also plated some glowing cells on amp plates (my first at home experiment) and I finally got them to glow. It was a great feeling when I first saw them. Now If I were to put these plates in the freezer to keep for a while, how long do you think I could save them for later use?

Also, I got an email back from the supplier of the gram-positive glowing plasmid and they said that there is information on the plasmid map on the site, he wasn't too clear about it. He also added that it is about $561 US for the plasmid, not very cheap.  

On Monday, July 30, 2012 11:18:48 AM UTC-4, Mega wrote:

Just wanted to tell,

I plated the glowing colis (from my bio fridge) on new plates, and they were much brighter than the original ones. Maybe because of less satellite colonies because of fresh amp ?
They were even so bright that you could see their glowing in a semi-dark room (10 o clock PM, streetlights illuminating the room weakly) without adapting your eyes to it.




Am Sonntag, 22. Juli 2012 01:15:33 UTC+2 schrieb Chowe:

Hello! Im very new to DIYbio and I am looking to do an exciting first project. I work in a genetics lab so I have experience with most techniques. I have had many ideas for projects, that I happily found out most people have had the same ones too (glowing plants, glowing yogurt) but I thought a lava lamp would be the simplest way. I am wondering how you guys would go about completing this. I saw that cambridge igem team made a lava-lamp, seen here http://www.youtube.com/watch?v=tUFscEVK5Ks I would like to make something just like that. I would prefer if I didn't have to induce it or if I did it would be through oxygen like the cambridge lava-lamp. It would be cool to have in my room to light it up at night. As with the glowing plants and yogurt they seem to be a lot harder and it seems like not many people have been successful so I wanted to choose a project that I would be more likely to succed. Thanks!

Corey

That is great to hear! I have also plated some glowing cells on amp plates (my first at home experiment) and I finally got them to glow. It was a great feeling when I first saw them. Now If I were to put these plates in the freezer to keep for a while, how long do you think I could save them for later use?

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Oh, that is *very* clever. My apologies, I was only running off what I
saw in the email thread! :)

On 31/07/12 22:04, Nathan McCorkle wrote:
> On Tue, Jul 31, 2012 at 4:59 PM, Cathal Garvey <cathalgarvey@gmail.com> wrote:
>> IMHO, Sugar would make a poor substrate for cell culture. The osmotic
>> pressure of growing on sugar would be huge; that's one reason why it's
>> traditional to add so much sugar to preserves like jam.
>>
>
> Cathal, they don't use the sugar to grow on, they print a mold with
> the sugar, then add gelatin + cells and pour it over the mold. Once
> the gelatin hardens, the mass is rinsed in an orbital shaker with
> media or buffer (not sure which) to dissolve the sugar, then that
> liquid is replaced with media. In Jordan's case, the mold looks like
> vasculature to aid perfusion later (he perfuses with orbital shaking,
> but I think he's also working on using a pump driven system).
>

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On Tue, Jul 31, 2012 at 4:59 PM, Cathal Garvey <cathalgarvey@gmail.com> wrote:
> IMHO, Sugar would make a poor substrate for cell culture. The osmotic
> pressure of growing on sugar would be huge; that's one reason why it's
> traditional to add so much sugar to preserves like jam.
>

Cathal, they don't use the sugar to grow on, they print a mold with
the sugar, then add gelatin + cells and pour it over the mold. Once
the gelatin hardens, the mass is rinsed in an orbital shaker with
media or buffer (not sure which) to dissolve the sugar, then that
liquid is replaced with media. In Jordan's case, the mold looks like
vasculature to aid perfusion later (he perfuses with orbital shaking,
but I think he's also working on using a pump driven system).

--
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Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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IMHO, Sugar would make a poor substrate for cell culture. The osmotic
pressure of growing on sugar would be huge; that's one reason why it's
traditional to add so much sugar to preserves like jam.

On 30/07/12 14:07, Jordan Miller wrote:
> here's our sugar extruder:
> http://blog.reprap.org/2012/07/on-challenge-of-3d-printing-sugar-for.html
>
> cheers,
> jordan
>
>
>
> On Jul 30, 2012, at 1:08 AM, "Jeremy[biohak.net]" <jeremya11z@gmail.com>
> wrote:
>
> No I do not. I wish I did. I don't really have access to the type of
> equipment necessary to accomplish that. I am interested though in maybe
> converting a rep rap to do sugar extrusions. Are the home cnc techniques
> that could work for something like this?
>
> On Thursday, July 26, 2012 8:50:59 AM UTC-5, Petfixer71 wrote:
>>
>> Do you have Any experience with cell culture on extracellular matrix??
>>
>>
>>
>> On Monday, July 23, 2012 3:07:40 AM UTC-4, Jeremy[biohak.net] wrote:
>>>
>>> Here's <http://www.biohak.net/node/12>here's an article I posted up on
>>> my blog. Apparently researchers were able to produce this jellyfish by
>>> grown muscle cells along the outer surface of a polymer mold.
>>>
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For *lab strain* E.coli, my understanding is that they're generally
unregulated unless they're GMO.

At least in Ireland for example, there is a legal framework regulating
Class 2+ bacteria for health and safety reasons. E.coli is in the list,
but with a footnote stating that domesticated lab strains are exempt.

To send E.coli in the post, send a "stab": create a little test tube of
slanted agar, then stab an inoculating loop of E.coli into the agar. You
can cap the tube (E.coli are facultative anaerobes, they won't need air)
and send it in a padded envelope. For added surety, alcohol-spray a
thick plastic ziploc bag, let it dry, put the tube in that before
mailing though. That way, if the tube breaks, the fragments will
possibly still be in a sanitised environment for recovery on the other end.

Label the letter "Non-hazardous biological sample": it's good manners
and could avoid trouble if customs at either end take exception to test
tubes of unidentified bacteria. Ideally, include a little letter slip in
the envelope describing the strain in case customs decide it's worth
opening. I've never heard of one being opened though; labelling it is
pretty "transparent". Postal service workers in most countries haven't
caught the "OMG BIOTERRORISTS" bug yet, it seems. :)

On 31/07/12 16:54, Andreas Sturm wrote:
> Maybe I can try to send you a lab strain from our university....
>
> But I fear, without them frozen they won't survive until the letter'd have
> arrived... Anyway, is it legal to send (genetically unmodified) Colis via
> mail??
>

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This article might be of interest to some DIYbio'ers who are also interested in life extension experiments.

230nm is used for phenol and related organic detection, it doesn't factor into the 260:280 number

And the ''no delivery to residental adresses'' is *often* not meant so seariously... I'd just  try it  ;) 
Don't mention you're doing diy bio, just write it as you were a lab and  would every day try out samples from companies.


Worst case: they don't answer your mail or  even worse, some conservative may tell you you shouldn't do experiments at home (one of the most stupid attitudes ever!). So you just don't answer back and never mind :)
 

Maybe I can try to send you a lab strain from our university....

But I fear, without them frozen they won't survive until the letter'd have arrived... Anyway, is it legal to send (genetically unmodified) Colis via mail?? 

Microinjection?

http://www.youtube.com/watch?v=h-Bfc1GPWpE

Looks cool... But, it will be hard to build it in a diy way... 

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Not so much complicated as extremely difficult. You need a
micromanipulator and microneedle for one thing, and then you need the
manual skill to use one effectively. Finally, you need lots and lots of
insect ova! :)

This is routine for making transgenic fruit flies, for example. I was
interested in trying it with sea monkies (Artemia salina) for a while
until I discovered how hard it is to get microneedles/micromanipulators.
I'm leaving it for later!

On 30/07/12 15:56, Andreas Sturm wrote:
>> theoretically you could also do somatic cell nuclear transfer (SCNT)
>>from a modified cell to an embryo
>
>
> Of course, but this seems even more complicated ;)
>
>
>
> 2012/7/30 Nathan McCorkle <nmz787@gmail.com>
>
>> yes you want to hit the germline to get expression in the whole
>> organism (or at least to get the new DNA in the whole organism, you
>> may add DNA that only expresses in one body area)
>>
>> theoretically you could also do somatic cell nuclear transfer (SCNT)
>> from a modified cell to an embryo
>>
>> On Sun, Jul 29, 2012 at 8:25 AM, Mega <masterstorm123@gmail.com> wrote:
>>> Hey,
>>>
>>> I read that it would be quite easy to add genes to insect cells using the
>>> piggyBac transposon. Frequently used for recombinant protein expression
>> in
>>> insect cell lines.
>>> So I was wondering how you could get an insect out of those cells. Do you
>>> have to transfect insect sperm? Insects are not plants, so you probably
>>> can't use any cell for reproducing an entire organism?
>>>
>>>
>>>
>>>
>>>
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>>>
>>
>>
>>
>> --
>> Nathan McCorkle
>> Rochester Institute of Technology
>> College of Science, Biotechnology/Bioinformatics
>>
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>>
>

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