Science

On 04/30/2016 09:29 AM, William Beeson wrote:
> I will test transformations on E.coli and Neurospora crassa. I've had interest in a low cost electroporator for months. I think
> the path to validation would be to show effectiveness with E. coli first (a much easier organism to transform). Then transition
> to fungal cells which have much lower transformation efficiency. If you have the hardware expertise to build a device with
> pre-programmed settings (say an E. coli and a fungal cells setting) I can do the lab work and prepare a data package for the product.
>
> -Will
>
> On Saturday, April 30, 2016 at 9:54:35 AM UTC-4, John Griessen wrote:
>
> On 04/29/2016 07:33 PM, William Beeson wrote:
> > If anyone has ideas for how to get this to work well please post or share the ideas. The amount of electrical engineering
> > involved in this is beyond my skill level.

How fast are you wanting this? I'm in the middle of another project.
Anyone else interested in early testing of prototypes? These will be "junk prototypes".
Similar to DIY art assemblages, but a little neater, and documented so a product
could be made from them by a few revision iterations.

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On Apr 29, 2016 10:48 PM, "BraveScience" <bravescience@gmail.com> wrote:
>
> Not all people have the knowledge, time, equipment and skills to isolate mycelium through clonal reproduction.

That is what the internet/books and friends/mentors at for.

> Giving them the chance to receive and use ready-to-use material doesn't sound a crazy ripoff but a good opportunity to start correctly.
>
> Moreover have you ever tried to isolate tissue from mushroom cap?

Yes, you pressure-cook some agar, get a few pressure-cooked knives/razors/scalpels, then wipe a mushroom with alcohol and maybe dip it in a bleach + sterile water solution, rinse well with sterile water, then shave layers off the mushroom, for each cut alcohol dipping and flame sterilizing the cutting tool. If it it really dirty, switch cutting tools. When you get a nice clean chunk exposed, place onto agar. Leave in kitchen cabinet for a few days and you will see mycelium, after a few months untouched it is not uncommon to get a small micro/mini mushroom.

> Much easier to have some already growing mothefucking thick piece of mycelium.

If you can't handle what I described above, there's probably not too much more someone would be able to do with a plate of mycelium, other than look at it die or possibly fruit. But I guess maybe there's a demand quickly available generic mycelium... I've never seen it sold like that before anywhere, so hard to imagine a use case.

On 04/29/2016 07:33 PM, William Beeson wrote:
> If anyone has ideas for how to get this to work well please post or share the ideas. The amount of electrical engineering
> involved in this is beyond my skill level.

Will you commit to some testing? Maybe 4 times on the same filamentous fungal cells? Over weeks with little
test circuit boards in the mail?

You're only interested in having this yesterday, so as soon as it works for you, the testing stops?
If it doesn't get going in a week, you're out?

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Not all people have the knowledge, time, equipment and skills to isolate mycelium through clonal reproduction.

Giving them the chance to receive and use ready-to-use material doesn't sound a crazy ripoff but a good opportunity to start correctly.

Moreover have you ever tried to isolate tissue from mushroom cap?
Much easier to have some already growing mothefucking thick piece of mycelium.

Cheers
Fede

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On 04/29/2016 02:23 PM, Nathan McCorkle wrote:
> Now thinking back to those discussions, there was a lot of concern
> with isolation. Would a wireless microcontroller obviate most/all
> concerns? Obviously isolation would still be good to protect the
> microcontroller... but assuming isolation development time cost more
> than a few $2 replacement ESP8266... well, does that assumption hold
> true?

The isolation that is needed is for safety, not economy, so no you can't skip it
and ship a product and not get bad karma, sued, end up way worse off than Willie Nelson
when his financial manager set him up with "tax shelters"...etc.

Anyone on this list interested in doing some of the leg work with me?
for a product with an oshwa.org pedigree as a kit, and same thing assembled and ETL tested
for North America/Europe lab equipment level of safety. For lab equipment your lawsuit
for product harm damages fails if you stick your tongue in it, but not for household stuff
'where babies rule'.


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I used a biorad micropulser at school, mainly for E.coli but it had a
yeast setting too, and maybe some things I don't remember. It wasn't
as big as the ones I see on ebay now.

On Fri, Apr 29, 2016 at 2:09 PM, Koeng <koeng101@gmail.com> wrote:
> Hey
>
> I have some organisms I would like to transform, and frankly,
> electroporation just works better. I would love to use an open source one,
> but that isn't happening very soon, so I have been considering for a while
> getting a used one off ebay. Does anyone have experience with a good pulse
> chamber + power supply combo? I would like to know what has been worked on
> (like perhaps the invitrogen electroporator ii) before I commit to a
> purchase.
>
> -Koeng
>
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Now thinking back to those discussions, there was a lot of concern
with isolation. Would a wireless microcontroller obviate most/all
concerns? Obviously isolation would still be good to protect the
microcontroller... but assuming isolation development time cost more
than a few $2 replacement ESP8266... well, does that assumption hold
true?

On Fri, Apr 29, 2016 at 9:54 AM, John Griessen <john@industromatic.com> wrote:
> On 04/28/2016 02:00 AM, Patrik D'haeseleer wrote:
>> Given how dirt cheap flyback transformers and other high voltage gear are
>> on my favorite online discount site
>> <http://www.goldmine-elec-products.com/products.asp?dept=1480>, perhaps
>> that might be a better way to go.
>>
>
> I have a back burner project to develop that. There are so few asking for it
> though, so I work on another project
> to generate income to have enough momentum to develop anything at all first.
>
> On 04/28/2016 03:53 AM, Marc Juul wrote:
>> Add an ESP8266 and you'd be able to connect via wifi+web to download and
>> view the conductance measurement and discharge graph for
>> use in your scientific article.
>>
>
> I'll be making a multi pulser flyback version so it can stretch a pulse,
> (with many peaks), out to 5ms or cook for 300 ms
> if desired. And then the power flow and impedance can be adjusted for
> different conductivity test cells. I'm a backer of
> the micropython on esp8266 and could easily use that one as a controller for
> this project. Are you going to commit time, and
> will you license it open hardware compatible?
>
> On 04/28/2016 03:12 PM, Nathan McCorkle wrote:
>> Is electroporation preferable? From the first time I used it, my first
>> summer internship, I was hooked. I thought "screw chemical
>> transformation" for the, in my opinion, more elegant, tuneable,
>> eco-friendly/greener-chemistry, faster, wider species coverage (you
>> just vary the pulse shape/voltage, maybe the salt content of the
>> buffer)... and higher efficiency to boot.
>>
>> Why isn't it more prominent? Because no one has made a cheaper or
>> open-source version yet.
>
> OK, so will you help me promote it with twitter, instagram, snap chat videos
> when the time comes?
> Who to sell to besides DIYers?
>
> There were some discussions I kept "best of" emails from before 2014 when my
> list server crashed
> without a backup I can find anymore. We were talking with a retired Oak
> Ridge nuclear engineer
> developing RF induction heaters using a car induction coil. It's all a
> little heavy and expensive compared
> to the flyback parts used in laser printers -- I have a box full of those to
> test with...
>
> The list has been up again since just after that crash if anyone wants to
> use it.
> http://lists.cibolo.us/mailman/listinfo/open_electroporator
> I think here on diybio@googlegroups.com is good for worthwhile
> milestones, and there have not really been any. Not much was lost from that
> crash.
> For saving old crusty details, there is anew list serve besides googlegroups
> by an original Yahoo.com
> businessman called groups.io -- it will have ads.
>
> John Griessen
>
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On 04/28/2016 02:00 AM, Patrik D'haeseleer wrote:
> Given how dirt cheap flyback transformers and other high voltage gear are on my favorite online discount site
> <http://www.goldmine-elec-products.com/products.asp?dept=1480>, perhaps that might be a better way to go.
>

I have a back burner project to develop that. There are so few asking for it though, so I work on another project
to generate income to have enough momentum to develop anything at all first.

On 04/28/2016 03:53 AM, Marc Juul wrote:
> Add an ESP8266 and you'd be able to connect via wifi+web to download and view the conductance measurement and discharge graph for
> use in your scientific article.
>

I'll be making a multi pulser flyback version so it can stretch a pulse, (with many peaks), out to 5ms or cook for 300 ms
if desired. And then the power flow and impedance can be adjusted for different conductivity test cells. I'm a backer of
the micropython on esp8266 and could easily use that one as a controller for this project. Are you going to commit time, and
will you license it open hardware compatible?

On 04/28/2016 03:12 PM, Nathan McCorkle wrote:
> Is electroporation preferable? From the first time I used it, my first
> summer internship, I was hooked. I thought "screw chemical
> transformation" for the, in my opinion, more elegant, tuneable,
> eco-friendly/greener-chemistry, faster, wider species coverage (you
> just vary the pulse shape/voltage, maybe the salt content of the
> buffer)... and higher efficiency to boot.
>
> Why isn't it more prominent? Because no one has made a cheaper or
> open-source version yet.

OK, so will you help me promote it with twitter, instagram, snap chat videos when the time comes?
Who to sell to besides DIYers?

There were some discussions I kept "best of" emails from before 2014 when my list server crashed
without a backup I can find anymore. We were talking with a retired Oak Ridge nuclear engineer
developing RF induction heaters using a car induction coil. It's all a little heavy and expensive compared
to the flyback parts used in laser printers -- I have a box full of those to test with...

The list has been up again since just after that crash if anyone wants to use it.
http://lists.cibolo.us/mailman/listinfo/open_electroporator
I think here on diybio@googlegroups.com is good for worthwhile
milestones, and there have not really been any. Not much was lost from that crash.
For saving old crusty details, there is anew list serve besides googlegroups by an original Yahoo.com
businessman called groups.io -- it will have ads.

John Griessen

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Or there is Theranos, where unverified data and grandios claims and press releases leads to a 9 billion dollar valuation evaporating. 

Ok, fair points. I'd direct criticism at the root problem, in that case: if "mycelium" is too nonspecific then it needs to be more specific, rather than cheaper but still vague. :)

On Thu, Apr 28, 2016 at 11:43 PM, Cathal (Phone)
<cathalgarvey@cathalgarvey.me> wrote:
> Don't do that, please. People devaluing one another's work to the
> lowest-common-denominator is the kind of peer anti-support that keeps
> everyone labouring in obscurity.
>
> Leaving asode the fact that a brick-and-mortar biohacking shop is costly
> (but it apparently exists!), there are significant advantages to buying a
> strain for the paltry cost of €30: strain identity can be known, sterility
> and purity guaranteed, and you're standardised with everyone else who bought
> that product and can avail of a community for support.
>
> I recall getting this crap when I was planning to sell an open source
> plasmid *customised for DIYbio* for around €45: cheaper than pUC18 at most
> vendors, but I distinctly remembering someone accusing me of profiteering
> for not selling at $1. As if years of work designing and testing had nothing
> to do with making the thing and only the costs of fermentation mattered. I
> was even threatened that I'd be deliberately undercut and pushed out of
> business.

I recognize a significant difference between the IndieBB plasmid and
"mycelium". It does not have any strain listed, and if someone really
wants "mycelium" they can get it for 100X cheaper, from common sources
including most rotting lignocellulosic material (unlike custom
synthetic engineered DNA) or for a GRAS option, a grocery store. I
didn't criticize the labelled strains they had which were apparently
collected to achieve a wide-ranging color palette from a single
vendor.

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Don't do that, please. People devaluing one another's work to the lowest-common-denominator is the kind of peer anti-support that keeps everyone labouring in obscurity.

Leaving asode the fact that a brick-and-mortar biohacking shop is costly (but it apparently exists!), there are significant advantages to buying a strain for the paltry cost of €30: strain identity can be known, sterility and purity guaranteed, and you're standardised with everyone else who bought that product and can avail of a community for support.

I recall getting this crap when I was planning to sell an open source plasmid *customised for DIYbio* for around €45: cheaper than pUC18 at most vendors, but I distinctly remembering someone accusing me of profiteering for not selling at $1. As if years of work designing and testing had nothing to do with making the thing and only the costs of fermentation mattered. I was even threatened that I'd be deliberately undercut and pushed out of business.

I hope Petshop.bio works out and I'm confident that the prices won't be the reason it doesn't. More likely the domain name will get seized by the organic association who own .bio. ;)