Re: [DIYbio] Isolate dNTP

It will be hard as the 'TP' (triphosphate) is a bit unstable (it
carries the high energy to fuel condensation reactions, i.e.
polymerization), or because most nucleotides in cell lysis solution
could be due to DNA degradation by DNases. You likely would need to
find a protocol to isolate mono or diphosphate nucleotides, then hit
them with a ATP and nucleotide kinase to 'recharge' them.

http://www.jbc.org/content/244/20/5489.full.pdf
http://pubs.acs.org/doi/abs/10.1021/bi00814a011

https://www.neb.com/products/m0201-t4-polynucleotide-kinase
https://www.neb.com/products/m0236-t4-polynucleotide-kinase-3-phosphatase-minus


This is a great project to think about, if you can determine an easy
method for purifying lots of material that can be converted with
kinase to dNTPs then we'd have a cheap DIY source for PCR and maybe
green chemistry DNA synthesis reactions.

On Thu, Feb 13, 2014 at 8:52 AM, Antonio Zafra <azafra93@gmail.com> wrote:
> I have an important question, how can I isolate dNTP? I think they are basic
> for using a PCR system.
>
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--
-Nathan

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