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role of water in PCR

PCR water

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#1 Shrubal

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Posted 06 December 2012 - 09:00 AM

Hello!

I was wondering if anyone could explain the specific role of water in a PCR reaction. I was wondering if the addition of water could be completely eliminated and instead compensated by adding more template if I had a very low concentration of template.

Thanks!

#2 Trof

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Posted 06 December 2012 - 09:19 AM

Depends whether you have your template disolved in water or in yogurt.

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#3 hobglobin

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Posted 06 December 2012 - 01:49 PM

Water is the solvent you use to get the final concentrations of the reagents in the desired volume...if you use more diluted DNA or other reagents (Mg2+, primers, buffer) you might end up that you don't need to add any additional water, but that's by chance then, as you cannot change their volume (except DNA often-times).
Anyway there's no need to add additional water as the water can come from any other source, but surely it's much easier to use it to get the desired volume.

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#4 mightymus

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Posted 18 December 2012 - 10:27 AM

Even I was having the same problem of low tempet.Try to evoporate the solvent (water or any buffer your dna is suspended) by heating the tube at around 55-60 C and increase the concentration of your dna. I did it a couple of times and it worked perfectly for my qpcr (:. By this way you retain the same final volume as you used to do before.

PS: make sure that the cap is open when heating ;)





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