dNTPs or deoxynucleotide triphosphates are the “building blocks” for DNA. Purity and stability of dNTPs are two of the essential factors to achieve a successful PCR. The use of a highly purified dNTP preparation is particularly recommended for sensitive techniques such as long-range PCR, RT-PCR, multiplex PCR, mutagenesis experiments and real-time applications. The purity of dNTPs is also important when the starting amount of template is minimal.

The standard concentration of a dNTP for PCR reactions is 0.2 mM. If the starting stock is a 25 mM solution of each dNTP in a 100mM dNTP Mix, you need to add 0.5 µL to a 50 µL standard PCR reaction.

It depends. You probably can increase the DNA yield, but you will have to optimize the complete PCR reaction, adjust the buffer, the Mg2+ and so on. It is not a matter pertaining only to nucleotides.

Yes. The sodium salt dNTP products are a standard grade and so we recommend these dNTPs for PCR. For more sophisticated reactions such as amplification of long templates, real-time PCR and NGS, we recommend lithium salt dNTP products.

Yes. The optimal pH for storage of nucleotides is from pH 7.5-8.2 (pH at 20°C). An acidic pH will cause hydrolysis of dNTPs (deoxynucleotide triphosphates) to dNDPs (deoxynucleotide diphosphates) and dNMPs (deoxynucleotide monophosphates), rendering them less suitable for PCR applications. During freezing/thawing cycles, the pH of the dNTP solutions can differ from the pH at 20°C. Sodium salts are temperature sensitive, so care needs to be taken when repeatedly frozen and thawed and sodium salt dNTPs are better aliquoted into smaller volumes and kept frozen in order to extend their shelf life.

The enzymatic synthesis of dNTPs uses highly specific enzymatic systems which eliminate impurities and PCR inhibitors, such as modified nucleotides, PPi and deoxynucleoside tetraphosphates. PCR reactions are impeded by the presence of contaminants resulting from chemical manufacturing processes, such as traces of dNDPs, pyrophosphates or other ionic species (e.g. acetate). Such contamination may lead to poor yields or to no PCR product at all. Unless thoroughly purified, chemically synthesized dNTPs often contain deoxynucleoside tetraphosphates which are powerful PCR inhibitors. Chemical synthesis can also lead to deamination and other nucleotide modifications whereas enzymatic synthesis of dNTPs bypasses these risks.

The concentration of our dNTP Mix are total – 100 mM dNTP Mix is made up of 25 mM of each dNTP (dATP, dCTP, dGTP and dTTP).

We recommend that you dilute your dNTPs using molecular biology or PCR grade water.

Yes, all of our dNTPs are glycerol-free and so can be lyophilized.