dNTPs or deoxynucleotide triphosphates are the “building blocks” for DNA. The 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 each dNTP in a PCR reaction is 0.2 mM. If the starting stock is a 100 mM solution of each dNTP, you need to add 0.1 µL of each nucleotide to a 50 µL standard PCR reaction. Since this is not convenient, it is recommended to prepare mixes: If the 100 mM dNTP stock solutions are mixed in equimolar amounts, the concentration of the mix will be 100 mM total or 25 mM of each nucleotide. From a 100x stock, you need to add 0.5 µL to a 50 µL reaction. Meridian also offers more diluted mixes of 40mM total (10mM each), which is a 50x stock solution, and 10 mM total (2.5 mM each) 10x working stock.

It depends. You probably can increase the DNA yield but you will have to optimize the complete PCR reaction, adjust the buffer, the Mg^2+, and so on. It is not a matter pertaining only to nucleotides.

是的。 对于复杂的反应，如长模板扩增和实时PCR, dNTPs的质量尤为重要。

是的。 储存核苷酸的最佳pH值为pH 7.5-8.2 (20°C)。 酸性pH会导致dNTPs水解成dNDPs和dNMPs，不太适合PCR应用。 在冻融循环期间，dNTP溶液的pH值可能与20°C时的pH值不同。 锂盐溶液的pH值不像钠盐溶液那样依赖于温度，因此在使用锂盐的地方，当dNTP反复冻融时，pH值不会发生剧烈变化。 因此锂盐dNTP更稳定，比钠盐有更长的保质期。

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, pyrophosphate, 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.

Reference was made earlier to the greater solubility of dNTPs in lithium salts than in sodium salts. Also, dNTPs presented in lithium salts are more resistant to repeated freezing and thawing than those presented in sodium salts. Furthermore, they remain sterile during the entire storage period (the lithium-ion has been shown to have significant bacteriostatic activity towards various microorganisms). Finally, using lithium-salt nucleotide preparations reduces salt-induced artifacts and increases the legibility of sequencing gels. Lithium salts are highly suited to PCR sequencing and labeling applications.

我们的dNTP混合物的所有浓度都是合计的，例如，我们的100mMdNTP混合是由每个dNTP (dATP, dCTP, dGTP和dTTP)的25mM组成的。

当dUTP代替或与dTTP一起使用时，所得到的PCR产物是尿嘧啶DNA糖基化酶(UDGase)的合适底物，这允许用户在开始当前的扩增之前完全摧毁任何来自先前PCR反应的污染DNA。 我们提供dUTP作为独立产品和dUTP Mix的一部分。

我们建议您使用分子生物级别或PCR级水稀释您的dNTPs。