Electroporator purchase help / advice sought

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Tom De Medts

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Mar 5, 2020, 12:49:53 PM3/5/20
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Dear DIY Bio community,

I seek your help in identifying a used, fully functional bacterial electroporator for garage lab use.

I've found a few on Ebay, but some of them are in the price range of $1000 - $3000, which is almost an order of magnitude more than my budget, which is ~ $300 - $500 including tax + shipping.

Often times, I find the setup includes, if it is BioRad,
the gene pulser, 
the capacitance extender, and 
the pulse controller, 
but missing the shocking chamber. I wonder if the cuvette chamber is the first item in the setup to breakdown? To make things worse, the cuvette chamber aka shock chamber costs > $500 just by itself.

So I wonder whether there are secret / hidden / obscure sellers, incuding but notl imited to Ebay, and LabX.com etc, from where I can buy an electroporator (BioRad preferred, but not mandatory) that is fully functional for bacterial gene transfer (not interested in eukaryotes).

Thanks, and some links from my online research are listed below.

Best,
Tom

JUST CHAMBER
https://www.ebay.com/itm/Biorad-MicroPulser-Electropcrator-Shock-Chamber-Assy/143546859131?_trksid=p2485497.m4902.l9144

FULL SETUP
https://www.ebay.com/itm/BIO-RAD-Gene-Pulser-Pulse-Controller-Capacitance-Extender-plus-Chamber-shock-qz/193210236659?hash=item2cfc3a3ef3:g:c7UAAOSw7oldztU1&autorefresh=true
https://www.ebay.com/itm/BIO-RAD-Gene-Pulser-II-Pulse-Controller-Capacitance-Extender-plus-Chamber-shock/153804889929?hash=item23cf7c5f49:g:NL0AAOSwgZpduPcM
https://www.ebay.com/itm/BIO-RAD-Gene-Pulser-II-Pulse-Controller-II-Capacitance-Extender-II-Chamber-4652/263745627533?_trksid=p2485497.m4902.l9144

PARTIAL SETUPS
https://www.ebay.com/itm/Bio-Rad-Pulse-Controller-Capacitance-Extender-and-Gene-Pulser/233318965856
https://www.ebay.com/itm/Bio-Rad-Pulse-Controller-Plus-with-MicroPulser-Shock-Chamber-II/274284018442

John Griessen

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Mar 5, 2020, 5:51:18 PM3/5/20
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On 3/5/20 11:49 AM, Tom De Medts wrote:
> Often times, I find the setup includes, if it is BioRad,
> the gene pulser,
> the capacitance extender, and
> the pulse controller,
> *but* missing the *shocking chamber*. I wonder if the cuvette chamber is the first item in the setup to breakdown? To make things
> worse, the cuvette chamber aka shock chamber costs > $500 just by itself.

I have a couple of bio-rad gene pulsers, (not the gene pulser II the first one), to compare to the culture shock electroporator
since classic electroporation recipes are done with a big cap discharging into a 30k Ohm load resistor with the cuvette in
parallel with that load, (and the loaded cuvette is usually going to be a good bit more resistive than the standard Bio_rad 30k
Ohms). These first ones have pulse extender units available on ebay for under $100. It does not come with a safety shrouded
cuvette holder, just two universally available banana jacks for the output volts and current. You can rig up anything springy on
a plastic base to do the cuvette holding.

I've been needing to do some characterization of them and send one out to Nathan, and the other could go to you...for $50+
shipping $25. After characterizing a gene pulser I, I'll be looking for a gene pulser II like you were seeing high prices for,
since more recipes use that, and culture shock will need some guides on how to get similar results to those many many old recipes
using gene pulser I & II... I think with an ebay search going I'll find a II, (with cuvette holder), for under $300 in a few months.

The culture shock electroporator is a prototype that can shock any old amount or conductivity of stuff tossed into a cuvette even
if under-filled to the correct field strength that will perfectly porate cells. It does so by measuring before it lets rip. It
also takes a time recording, (by being a digital oscilloscope), so you can fire off blasts to cuvettes willy nilly and if you
label them, figure it out later or delegate that figuring to someone else. If that sounds useful, email me for more info. It's
not ready to go in a normal lab setting yet -- no case, no pushbuttons -- but more on that before year end.

John Griessen

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Mar 5, 2020, 6:03:46 PM3/5/20
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On 3/5/20 11:49 AM, Tom De Medts wrote:

> pulse controller,
> *but* missing the *shocking chamber*.

I just looked at ebay and remember now it was the Gene Pulser Xcell that has the little pod for holding a cuvette with safety
shield. I'm going to want to get one of those, and they are going to be more $$, probably $600 with some waiting. The Xcell
models have a little programmability, but not as much as culture shock is going to have, (culture shock already has it, just not
easy-to-use lab-ready).

Nathan McCorkle

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Mar 5, 2020, 10:26:46 PM3/5/20
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Like John said, what you're calling the shock chamber is nothing more than a holder with spring loaded contacts for either side of the disposable* electroporation cuvettes. (* People have claimed getting many uses out of these by sterilizing with HCl or other obvious chemical means).

If you have a 3d printer or basic wood-shop level fabrication tools, you can easily rig something up. Just make sure it's easy/quick to remove the cuvette, since the time from shocking to adding fresh media needs to be minimal.

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Tom Hodder

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Mar 6, 2020, 6:44:30 AM3/6/20
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On Thu, Mar 5, 2020, 9:49 AM Tom De Medts <tdm...@gmail.com> wrote:
but missing the shocking chamber. I wonder if the cuvette chamber is the first item in the setup to breakdown?

In a similar situation, we received an Eppendorf multiporator which was missing the cuvette holder. We ended up improvising something on the laser cutter, and using some copper strip to make the connections to the cuvette and PSU











 

Tom De Medts

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Mar 9, 2020, 10:27:17 AM3/9/20
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Hi folks,

Thanks for sharing your experience, expertise and even offering to send / sell me your
fabricated shocking chamber. Much appreciated.

I am a biologist, and not a real DIYer. Since I am so used to university
based labs, I bit the bullet and purchased a used Bio-Rad electroporation setup
including the gene pulser, pulse controller, capacitance extender and the shocking chamber.

This was a $550 purchase, which is not small change for garage lab operators like me. So
I hope it works as it is expected to.

But in case this set up fails to work as expected, I'll be sure to ping you helpful
folks again.

Pierre Padilla - The Portoporator is super cool and interesting. My friend did contact
Dr. Daniel Gilbert in Germany, who suggested getting local help for fabrication.
This same friend also contacted Dr. Saad Bhamla at Georgia Tech, of ElectroPen fame,
and he also suggested getting local help. So connecting with local DIY talent is something
I need to pursue actively...
A few, select ElectroPen / Saad Bhamla links -
https://twitter.com/bhamlalab/status/1076331619743608832?lang=eu
https://twitter.com/bhamlalab/status/1076334209923534849?lang=en
https://www.youtube.com/watch?v=yPwn0nexCtU
https://www.biorxiv.org/content/10.1101/448977v1

John Griessen - I don't think I have the skill set to use your custom shocking chamber.
But should the BioRad setup arrives soon not work as expected, I will take you up on your
offer.

Nathan McCorkle - Agreed, I need to establish connections with the local community college,
where I am sure undergrad kids have experience and expertise with additive manufacturing.
Thanks also for the warning about minimizing time between shock and medium addition.

Cheers,
Tom

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Nathan McCorkle

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Mar 10, 2020, 2:36:49 PM3/10/20
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Please note that electronic lighter piezoelectric elements can vary significantly in voltage output, by at least 10x depending on a particular given lighter source. It just means you need more testing to classify a piezo element before one of those lighter based circuits will work effectively. 

Tom De Medts

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Mar 15, 2020, 3:46:48 PM3/15/20
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Hi folks,

The BIO Rad gene Pulser I electroporator arrived from Ebay today.

Can someone share a wiring diagram (even if hand drawn) for a setup containing the modules of the BIO RAD Gene Pulser I Electroporator, consisting of 
  1. Gene Pulser (bottom most)
  2. Pulse Modulator (mid tier)
  3. Capacitance Extender (top most module)
as shown from 4 annotated pics below of the 4 different sides of the stacked modules, please?

I am not entirely sure how the wires from each module goes to the next one, hence this request and the accompanying pics. Thank you, in advance.

Do I even need the pulse modulator and the capacitance extender, if all that I intend to use this for, is electoporation of E. coli or Agrobacterium?

Cheers,
Tom

PS. You should be able to download and zoom into the pics, if you wish. Or open in a new tab, and expand...

Front_setup.jpg Right_setup.jpg Back_setup.jpg Left_setup.jpg




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Tom Knight

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Mar 15, 2020, 7:13:42 PM3/15/20
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I’d recommend that you unplug the capacitance extender and pulse controller and put them in the closet until you think you need them. The base unit is fine for almost all work. If you are doing bacterial samples, aim for a time constant of 5 ms with samples loaded. The base unit puts a 200 ohm resistor in parallel with the sample chamber, so you can try this out with no sample loaded. Adjust the capacitor to give a 5 ms pulse. If you samples are high resistance, you should not need to change anything. For E. coli, try 1.8 KV with a 2 mm cuvette, possibly up to 2.2 KV. For 1 mm cuvette, try 1.2 KV or so.


On Mar 15, 2020, at 3:46 PM, Tom De Medts <tdm...@gmail.com> wrote:

Hi folks,

The BIO Rad gene Pulser I electroporator arrived from Ebay today.

Can someone share a wiring diagram (even if hand drawn) for a setup containing the modules of the BIO RAD Gene Pulser I Electroporator, consisting of 
  1. Gene Pulser (bottom most)
  2. Pulse Modulator (mid tier)
  3. Capacitance Extender (top most module)
as shown from 4 annotated pics below of the 4 different sides of the stacked modules, please?

I am not entirely sure how the wires from each module goes to the next one, hence this request and the accompanying pics. Thank you, in advance.

Do I even need the pulse modulator and the capacitance extender, if all that I intend to use this for, is electoporation of E. coli or Agrobacterium?

Cheers,
Tom

PS. You should be able to download and zoom into the pics, if you wish. Or open in a new tab, and expand...

<Front_setup.jpg> <Right_setup.jpg> <Back_setup.jpg> <Left_setup.jpg>




On Tue, Mar 10, 2020 at 2:36 PM Nathan McCorkle <nmz...@gmail.com> wrote:

Please note that electronic lighter piezoelectric elements can vary significantly in voltage output, by at least 10x depending on a particular given lighter source. It just means you need more testing to classify a piezo element before one of those lighter based circuits will work effectively. 

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