dNTP and MgCl

Description

A typical amplification reaction includes target DNA, a thermostable DNA polymerase, two oligonucleotide primers, deoxynucleotide triphosphates (dNTPs), reaction buffer and magnesium. dNTP Mix is a solution containing sodium salts of dATP, dCTP, dGTP, and dTTP, each at 10 mM in water at pH 7.5 while the total concentration of nucleotides is 40 mM. Template DNA concentration, chelating agents present in the sample (e.g., EDTA or citrate), dNTP concentration and the presence of proteins all can affect the amount of free magnesium in the reaction. In the absence of adequate free magnesium, Taq DNA Polymerase is inactive. Excess free magnesium can reduce enzyme fidelity and increase the level of nonspecific amplification. The performance of Pfu DNA Polymerase seems depend less on magnesium concentration, but when optimization is required, the optimal concentration is usually in the range of 2-6 mM. Using dNTP during the extension phase provides single bases ready to go into DNA and double it, similar like building blocks. dNTPs are preferred nucleotide substrates in producing either homo- or heteropolymeric tails. The dNTPs are the artificial nucleotides used in the PCR to synthesize new DNA strands, such as DNA replication. Each dNTP is made up of a phosphate group, a deoxyribose sugar and a nitrogenous base. dATP and dGTP make up the purines which have a double ring structure. dTTP and dCTP make up the pyrimidines which have a single ring structure.

Features and Benefits:

• Simple protocol: reducing pipetting steps, reduce the possibility of contamination.
• Inhibit subsequent amplifications: incorporate dUTP into PCR products and treated by uracil-N-glycosylase.
• Highly pure: greater than 99% triphosphate content and provided at a concentration of 100 mM in water at pH 7.5 which is allowing highly consistent results.
• Preventing false positive: Uracil-N-glycosylase can be added to a PCR premix to excise uracil from any contaminating PCR product.
• Optimal: PCR-tested deoxynucleotides ensure optimal performance with all Promega amplification enzymes. Addition of GoTaq® Flexi Reaction Buffers with Magnesium Chloride Solution can optimize PCR Reaction.
• Convenience: supplied at a convenient concentration (100 mM in water) for versatile use in PCR and other applications.