Cheapest and easiest way to do PCR at home?

[Avery Ashley]

What's your setup?

How much did it cost?

Any recommendations for someone who desperately wants a PCR machine but can't afford one and doesn't have the skills to delve into DIY electronics?

[Avery louie]

honestly, I would try ebay.  OpenPCR is pretty expensive (~600?), for that kind of money you can get an ebay/refub machine and not have any of the stress.

Or you could kick it old school and "do it with sticks and rocks" as i like to say, aka use water baths and manually move the tubes.

Roughly where are you located?  Are there any local groups?

--A (the other Avery)

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[Avery Ashley]

I'm in the middle of nowhere, southeast North Carolina. I haven't officially checked, but I seriously doubt there's a group near enough me. Plus, I'm really a novice with all this stuff, I wouldn't be of any use to anyone for a while. I'd like to work it all out myself before I work with others.

I suppose you're right though, I can try and save three or four hundred in the next month or two.

[Avery louie]

I don't know, it may be a good idea to get another person involved.  It certainly helps to split the burden and if you get frustrated, it is good to have a friend.

--A

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[Nathan McCorkle]

You can try getting polymerase free samples that include control DNA
and nucleotides, but you'll have to buy agarose and some buffer to do
electrophoresis and check to see if the PCR worked...

If you called mobio and said you wanted to buy their cheapest agarose,
$88 for 50 grams (which is enough for 50-100 gels) I know they would
give you free samples to try out PCR (since they already offer most of
what you need for free samples)
http://www.mobio.com/electrophoresis-reagents/ultraclean-agarose-molecular-biology-grade.html

http://openwetware.org/wiki/DNA_separation_in_different_agarose_gels

You could do the thermo cycling with hot water and a thermometer in
styrofoam coffee cups (seriously)... so any automation in this process
is improvement!

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[Mega]

My proff told me that in former times they used wather baths :D A thermocycler is much more confortable though.

[ruphos]

Greatest efficiency is actually achieved through water baths. Ramp time is comparatively minimal when you are physically moving a tube between cold and warm bath. I thought about putting together a rotating rack with programmable step distances, but decided I was lazy and breadth of application was fairly small. If you can set up an environment that can easily regulate baths at several temps, it may still be worthwhile and likely rather simple to put together. Perhaps a project for our compatriots working on the most shoestring of budgets.

On Wed, Jan 23, 2013 at 1:03 AM, Mega <masters...@gmail.com> wrote:

My proff told me that in former times they used wather baths :D A thermocycler is much more confortable though.

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-- Friedrich Nietzsche

[Avery Ashley]

I'd be very interested in helping create a standardized water bath technique for the lower budgets. Anyone care to collaborate? I'm afraid I don't know enough about the process to do it alone, but I'd be willing to shell out the money for the parts.

Maybe we could make plans then provide them to the community for free?

On Tuesday, January 22, 2013 10:44:26 PM UTC-5, Avery Ashley wrote:

[Matt Lawes]

Probably disclosed in the original Cary Mullis / Cetus Corp patents for PCR. And a number of preceeding publications with DNA Polymerases.

Should speed up the job of protocolling.

>matt

Sent from my Verizon Wireless 4G LTE DROID

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[Koeng]

I would be interesting in helping as well, even though I have a working, good, programmable, thermocycler :)
As they said, ebay got the best prices. I got mine for 80 bucks and it works GREAT!

[Avery louie]

What model do you have? Many models go for similar prices.

--A

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[Mac Cowell]

How about two 5cm peltier junction plates, one big heatsink, a 5cm x 5cm x 1.5 cm milled block for holding tubes, and a simple chassis for sandwiching it all together?

Not sure what the power requirements are - would a 28 watts power supply (2 A 12 V) be too small for cycling the peltiers?  If the power supply was figured out, then an arduino and some power resistors could be used to control the peltiers.  Oh, and it'd need one or two thermocouples / thermisters.

Call it the CheapoPCR.  The BOM at least shouldn't cost more than $75.  Probably would have very slow ramping times... but hey, for $75 - $150, that's not bad.

Mac

On Wed, Jan 23, 2013 at 7:29 AM, fiaraz <fiara...@googlemail.com> wrote:

oh yep the convection cycler cost should be under $10 but only one vial at a time.

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[Sebastian S. Cocioba]

At one point it may be more economical to produce your own polymerase. I am not sure exactly at what point in house production beats commercial purchases. The lowest price I've seen is the econoTaq polymerase sold from sydlabs.com  which comes in a kit with buffer, dntps, and polymerase for $0.16 per reaction. IMO that is the dirt-est of cheap.

Sebastian S Cocioba

CEO & Founder

New York Botanics, LLC

Sent via Mobile E-Mail 

On Jan 25, 2013, at 7:04 AM, Adam Graham <grey....@gmail.com> wrote:

 Here is the original patent. Matt appears to be right that this does in fact contain basic protocols to carry out PCR. I haven't looked into it properly though.

Don't forget it costs money to obtain all the reagents. Not to mention you want to make sure your primers are right.

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[Dakota Hamill]

Sydlabs had great prices on everything but...never actually had the item.  I went to order Tris, EDTA, tips, tubes, and some sodium acetate from them, pretty common things you'd think they would have.  Got an email back saying they were out of everything we had ordered, never said when it would be in or made any effort to try to actually get them for us. 

I don't know if it's one dude outside Boston who drop ships stuff or what but, I wasn't very impressed.

Have you been able to get the Taq from them before?

[Sebastian S. Cocioba]

Yeah I bout the taq and it worked out fine. I but almost all my reagents from then. They do seem to have an issue with maintaining inventory from time to time but they are relatively new. I've been bargain hunting for reagents for a while now and they seem the cheapest if its in stock.

Sebastian S Cocioba

CEO & Founder

New York Botanics, LLC

Sent via Mobile E-Mail 

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[Dakota Hamill]

Alright good to know.  I'll give them another shot.

[biomiky]

Hello folks!

I'm a hobbist biology from Italy and currently unemployed for years. My dream is to build up a homemade "kitchen" molecular biology lab. Is there anyone who want to be involved in such a project?

My budjet is very low: 2000 to 7000 euros. 

Bye

Michele

[Cathal Garvey]

Hey biomiky,
You might like to join the DIYbio-eu list as well, as we'd have better
suggestions for suppliers to Italy. Just send an email to:
diybio-eu...@diybio.eu

2-7K is more than enough to set up a basic MolBio lab, if you're
willing to be very frugal. I set up the very basics of my lab for about
2K, though since then I've spent much more improving it. I have an old
spreadsheet lying around detailing the prices and suppliers that got me
there, I'll see if I can dig it up. So, don't be disheartened! :)

Slight problem for you might be that some of the suppliers won't like
shipping to Italy, as is the case with many Ebay sellers, but hopefully
that won't be a serious barrier. If it's important enough, I'm sure
someone on the lists would be happy to relay small packages your way..

best,
Cathal

[Mega [Andreas Sturm]]

Hi!

Where are you from?
Perhaps it would be great to start a community lab, if you find 10 people you can afford any machine :D

I've got friends in the Lazio region, maybe someone may be interested...

Best,
Andreas

[Koeng]

You can go.. very... very.. cheap. I have done hand PCR and a single pipette can do a lot. I have estimated I cloned some DNA for less then 300 bucks at the beginning (Leftover kit parts and minimal primers plus hand PCR and regeants from my bathroom). It was terrible. Don't go that cheap. I am so lucky it actually worked and it took over a month for a single DNA

-Koeng

[Cornelia]

You can also outsource your single reactions. No need to buy all the machines and reagents upfront. Check out this website: www.transcriptic.com
You can also do single , one-off reactions and pay per reaction. Just email them.

Of course it does take the fun of pipetting yourself, but at least you know that everything works.

Good luck!

[Koeng]

Their qPCR service actually looks quite interesting

[Michele Stefanoli]

Dear Cathal Garvey,
You did say:

"I have an old spreadsheet lying around detailing the prices and suppliers
that got me
there, I'll see if I can dig it up".

Could you gently provide this precious spreadsheet of suppliers?

Bye
-----Messaggio originale-----
From: Cathal Garvey
Sent: Wednesday, October 02, 2013 3:33 PM
To: diy...@googlegroups.com
Subject: Re: [DIYbio] Re: Cheapest and easiest way to do PCR at home?

[Sebastian Kraves]

check out www.minipcr.com.  Comes assembled, calibrated, and quality tested.

On Tuesday, January 22, 2013 10:44:26 PM UTC-5, Avery Ashley wrote:

[Sebastian Cocioba]

I don't mean to offend or step on toes but I am curious: what is the difference between Genefoo's personal pcr and the minipcr. Looks identical but one is a hundred dollars cheaper. I'm really looking into a small pcr machine but that cost can't be justified beyond niche market parameters. Cost of parts plus any reasonable profit margin can't equate to $800 unless the cost is being cranked up due to low demand, or MSRP based on other machines. I mean seriously, a peltier, custom micro, a PID program with thermocouple feedback should be the big cost driver. A custom machined the block may be pricey but not $800. Even OpenPCR shouldn't be that expensive. I really can't see a good way to justify the price. No offense.

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[Peter Thielen]

Interesting to note that Genefoo's website went down after your last post, and many of their products are now available on the minipcr website.  These are the same design, but I don't believe Genefoo was actually shipping completed units -- I anticipate that they have joined forces.  

Also of note, any mention of minipcr being open source has now disappeared from their website.  Pricing of individual components for assembling a personalPCR unit was in the $100-125 range last I checked, so the $800 figure has to involve paying people for development, testing, etc.  I don't fault them for the price hike, but I also don't see a lot of individuals making a purchase for a less functional thermocycler than can often be found in the used market for less money.  Support is key, though, so if they manage to make a business out of this, I'll be on the lookout for prices to drop.  

[Michele Stefanoli]

I' working on a pcr light bulb project i only need to experiment and assemble. Anybody did it? What do you think about it?

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[Otto Heringer]

Hi,

I'm from Brazil and we're planning to build a open DIYbio lab on our local hackerspace - and as you may see, I'm a newbie here too.

We're planning to build this guy here: http://www.instructables.com/id/Arduino-PCR-thermal-cycler-for-under-85/

OK, you will need a pro acocunt to download the file; and some parts seem to be hard to find. But it could be a nice and cheap project to try out.

[Michele Stefanoli]

What high power resistors nedded? Like some from an iron?

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[Otto Heringer]

They used this: http://br.mouser.com/ProductDetail/Vishay-Dale/RH050150R0FC02/?qs=oFi4/QsgZXYdykWTBf8Cwn8R2/shRA70

[Michele Stefanoli]

Nice. And for the aluminium dissipator? Thanks

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[Otto Heringer]

Although you cannot download the file on Instructables, you can see the steps for the building process online.
Take a look here for "step" about the aluminium block they've used:
http://www.instructables.com/id/Arduino-PCR-thermal-cycler-for-under-85/step3/The-aluminum-block/

This might be the hardest part to build because of the need of proper equipment for cutting.

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[Michele Stefanoli]

/products/amplification_pcr/product_overlay_content/global/lsr_ptc200_temperature_control.jpg

what TE stands for?
Thanks
Ms

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[biomiky]

Hi, 

I devised this machine that I want to validate. Tell me your opinion. Bye 

MS

[Otto Heringer]

Iron? Iron!? Did you mean iron clothes!?

haha It will be very interesting, please, DO IT! :)

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[Michele Stefanoli]

Yes. definitely! As soon as possible I gonna try and share some pics.

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[Nathan McCorkle]

Seems like the clothes iron would have a LOT of thermal mass, making
it harder/slower to cycle from hot to cold.

[Michele Stefanoli]

Ok. So thermal inertia is the issue. Would a 250W Ir lamp be the solution cooling up faster?

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[Michele Stefanoli]

I think that high power resistors are the best.

[Nathan McCorkle]

On Sat, Sep 20, 2014 at 4:34 AM, Michele Stefanoli
<micheles...@gmail.com> wrote:
> I think that high power resistors are the best.

A great and commonly available resistor with a fan is a hair drier.
Air also has the great property of low thermal mass.

[John Griessen]

On 09/20/2014 05:44 AM, Michele Stefanoli wrote:
> So thermal inertia is the issue. Would a 250W Ir lamp be the solution cooling up faster?

Many cyclers down the ages, (last twenty years), have used mechanical means to
switch hot cold quickly. They break the heat flow path by movement. A cam moves the thermoelectric
cooler heat sink out of contact with the samples-heater-block, then heaters turn on, heaters turn off,
cam moves back.

250 Watts? Depends on mass of samples/heat sinks/etc. Could easily be smaller. Could be a hair dryer
at 1000 Watts as well like Nathan suggests.

Air as the samples contact medium is the easiest way to switch heat flow back and forth if you ask me.
Think of "any kind of hot air source" blowing all the time, and a light weight door in a Y branch
in the air path to switch the air stream into the samples area or bypass it. Another ambient air
blower feeds another air path with Y branch flap valve to choose cool ambient air into samples
area or bypass. Next you just control the movement of these valves on the air ducts to
get thermal cycling. Use sensors at the heater box and at the samples zone
to tell how close to desired temperature you are. Preheated air flow needs to be fast
and wide swirling past your sample vials to get even and rapid heat transfer.

Recycle hot air that has either gone through the samples area or been bypassed around it to the intake
of your heater box to save energy. Let a separate control system regulate the heater to keep the
heater box exhaust at a reasonable temp you determine to get good heat flow rates.
Maybe the preheated air temp will be 140 deg C, maybe 118 deg C, maybe 180.

Testing will tell.

[Michele Stefanoli]

Thankful for your advices.
Ms

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[Sebastian Kraves]

Hey there,

miniPCR is on Kickstarter starting at $399

https://www.kickstarter.com/projects/minipcr/minipcr-a-dna-discovery-system-for-everyone

Thanks for checking it out.

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