Re: [DIYbio] Re: What's wrong with open source PCRs

Hi all,

@Ujjwal: that's a cool question. Personally, as it has pointed out above, connectivity is an issue. Even benchtop lab thermocyclers are kind of a pain to control smoothly. Actually they are horrible machines. Although I am part of the professional clients and my needs aren't always the DIYbioer's needs..

Price as well is challenging. There's a lot of technology to assemble and run properly and reliably. And price is very important for professional too.

All the suggestions so far pointed on making the system more efficient/cheaper and increased user experience (my point as well).

It's kind of remarkable, as well as happened for the fax, how thermocyclers didn't really change. They were horrible boxes and they still are. 

Like we weren't creative enough to come up with other systems that allow to heat/cool down some small amount of samples.

A good point was made by Jonathan: it's not too important to make a "cheaper" PCR machine by itself, but it's mandatory to reduce the amount of sample volume. What you can do in 50-100uL today you could easily scale down to 1-10uL. If you scale down to 1uL mix, well, it would be much much cheaper.

During colony PCR we usually use 10-20uL total volume. That means 5-10uL myTaq mix (all included). And it always works. I even used it for cloning, fuck those proofreading crappy polymerases. 

let's crunch some numbers: price tag is 104$ for 50uL reactions. That means 25uL master mix. Each reaction is 2$.

Going down by 10x means 5uL reactions would mean reaching 0.20$/reaction. Even in today's lab you can do quite a lot with 5 uL PCR volume if you reach a final yield of 100ng/uL. Enough to do plenty of cloning IF you avoid those horrible spin column/gel extraction kits... Gel extraction looses up to 90% of your stuff, especially if it >5kb.

Anyhow my point is size. Yet, and i know many will share this point, working with less than 10uL is a pain. 

If you work in a tube. 

Because pipetting isn't easy after the psychological barrier of <2uL and capillarity is a bitch.

Why don't we leave behind our backs those tubes? Come up with something new. 

Kind of muhammad story for molecular biology, if you cannot bring the heat to the sample, bring the sample to the heat. 

Samples could move across temperature gradients. You could get either low volume sizes and user friendly equipment.

Illumina and nanopore are already doing this. All sample preparations is ran on digital microfluidics biochips, although a patent fence as high as they sky is out there, and they perform an enriching amplification step previous to sequencing.

Best,

Fede

Il giorno giovedì 21 gennaio 2016 22:54:55 UTC+1, Ujjwal Thaakar ha scritto:

Hi,

I recently decided to design a PCR machine that's affordable since I can't either buy one or afford an OpenPCR. I wanted your views on OpenPCR, MiniPCR and the likes of others. Are these runaway successes? Did everyone really buy one for themselves? What were the issues you had with them? What does diybio need today the most and at what price point. 

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Thanks

Ujjwal

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