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Extremely desperate noob question: How do these PCR work?


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#1 FountainPenAddict

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Posted 22 March 2014 - 03:19 AM

Hello, I'm planing to make (http://www.popsci.co...04/gene-machine) or (http://www.instructables.com/id/Arduino-PCR-thermal-cycler-for-under-85/) and I'm wondering how they work? If they just replicate the DNA how is the DNA tested? How do you check it to see if a fruit has been genetically modified? 

 

Please help me. Thanks :)



#2 hobglobin

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Posted 22 March 2014 - 05:24 AM

as mentioned in the first link you need reagents (buffer including Mg2+, enzyme (taq polymerase), nucleotides, primers), no idea where you can get them as a private person. Finally you have to prepare (isolate) DNA, for this quite good protocols using household reagents exist.

And finally to make it visible usually gel electrophoresis (usually agarose gels (a polysaccharide matrix) in special gel chambers were DNA is separated by charge and size using an electrical field and then dyed and made visible e.g. by UV light. For this surely home made solutions (e.g. usig filter papers) exist too.

Not really sure if these devices will really work, especially the one in the first link seems to slow for a good working PCR, i.e. the system is too slow heating and especially cooling down again the water bath (professional thermal cyclers heat and cool at a rate of several degrees Celsius per second!). To have separate chambers with a fixed temperatures (three for denaturating, annealing and elogation) is much easier and reliable. Then you switch the tubes between chambers manually and have a fast and accurate temperature ramps too (the first professional machines worked similar). 

The second machine looks better but haven't looked in the details.


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...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

#3 Tabaluga

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Posted 22 March 2014 - 07:00 AM

Hello, I'm planing to make (http://www.popsci.co...04/gene-machine) or (http://www.instructa...r-for-under-85/) and I'm wondering how they work? If they just replicate the DNA how is the DNA tested? How do you check it to see if a fruit has been genetically modified? 

 

Please help me. Thanks smile.png

 

Interesting DIY websites, thanks for sharing !


Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#4 hobglobin

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Posted 22 March 2014 - 09:17 AM

 

Hello, I'm planing to make (http://www.popsci.co...04/gene-machine) or (http://www.instructa...r-for-under-85/) and I'm wondering how they work? If they just replicate the DNA how is the DNA tested? How do you check it to see if a fruit has been genetically modified? 

 

Please help me. Thanks smile.png

 

Interesting DIY websites, thanks for sharing !

 

the second site has the disadvantage that you have to pay for a "pro-account" to get more detailed information dry.png


One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

#5 El Crazy Xabi

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Posted 23 March 2014 - 04:43 PM

There is also a Google group called DIYBIO which also have a web http://diybio.org/



#6 FountainPenAddict

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Posted 25 March 2014 - 01:43 PM

 

 

Hello, I'm planing to make (http://www.popsci.co...04/gene-machine) or (http://www.instructa...r-for-under-85/) and I'm wondering how they work? If they just replicate the DNA how is the DNA tested? How do you check it to see if a fruit has been genetically modified? 

 

Please help me. Thanks smile.png

 

Interesting DIY websites, thanks for sharing !

 

the second site has the disadvantage that you have to pay for a "pro-account" to get more detailed information dry.png

 

You only need the pro account to download the PDF



#7 FountainPenAddict

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Posted 26 March 2014 - 10:56 PM

anyone else? 






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