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Deoxynucleotide Triphosphates (dNTPs)
Chemical structures of the four building blocks for nucleic acids, as included in the ReadiUse™ dNTP Mix Set *10 mM PCR Grade*.
Function of Deoxynucleotide Triphosphates (dNTPs)
Though it has been proposed that dNTP availability influences the progression of the cell cycle, the underlying mechanisms between dNTP levels and the cell cycle remain unclear. It is known, however, that strict maintenance of dNTP pools is critical for multiple cellular pathways, which means that both nucleotide synthesis and degradation are exquisitely fine-tuned within the body.
Imbalances in dNTPs are associated with genomic instability, tumor growth, and abnormal cellular development. Imbalances in dNTPs have also been shown to interfere with mitochondrial DNA (mtDNA) and mitochondrial health that could lead to related diseases like diabetes, obesity, or even cancer. To further underlie the importance of dNTPs for cell behavior, purine and pyrimidine metabolism (DPPM) disorders have also shown to profoundly affect cell metabolism. DPPM diseases can manifest in a number of ways including anemia, immunodeficiencies, mental and/or growth retardation, or serious adverse reactions to medication.
As fine-tuned dNTP levels are a vital key for healthy cellular homeostasis, over the years medicine has been expertly curated to combat different types of abnormal dNTP pathways. For example, in the treatment of several types of cancers common therapies are aimed at dysregulating abnormal dNTP synthesis and/or metabolism. Chemotherapies like methotrexate, hydroxyurea, and antimetabolite compounds like 5-fluorouracil generally work in tandem to limit the dNTP pool available for tumorous DNA synthesis, as well as trigger a cascade of events within the cell to result in cell cycle arrest.
Role in dNTPs in Polymerase Chain Reaction (PCR)
dNTPs are necessary components of PCR and, in fact, no amplified DNA products could be generated without them. dNTPs are used during the extension phase of PCR and provide nucleotide building blocks to the unzipped template strand of the DNA. By doing so, double stranded DNA is synthesized, which can continue to grow for the length of the template strand. dNTPs are generally added to PCR and RT-PCR mixes in equimolar amounts. However, it is ultimately important to think about the target DNA sequence to be amplified.
Note: If the DNA is mainly dATP/dTTP (AT ratio) or dGTP/dCTP (GC ratio), the amount of respective dNTPs in the PCR mix may be altered as necessary.
Some PCRs may benefit from highly purified “PCR-grade” dNTPs that are >99% pure, and free from contaminants like modified nucleotides or tetrapyrophosphate. Though they are more expensive, high-purity dNTPs may help successfully amplify products from samples that may contain very low copy numbers of target DNA.
In some PCR applications, one of the four dNTPs may also be replaced by an analog dNTP for downstream post-PCR processing applications. Common modified dNTP analogs include 7-deaza-DGTP, or a biotin-, fluorophore- or radioactively-labeled dNTP.
Product Ordering Information
Table 1. dNTP Conjugates
| Cat# ▲ ▼ | Product Name ▲ ▼ | Unit Size ▲ ▼ |
| 17014 | Biotin-11-dATP | 25 nmoles |
| 17015 | Biotin-11-dGTP *1 mM in Tris Buffer (pH 7.5)* | 25 nmoles |
| 17019 | Biotin-14-dCTP *1 mM in Tris Buffer (pH 7.5)* | 25 nmoles |
| 17035 | Fluorescein-12-dCTP *1 mM in Tris Buffer (pH 7.5)* | 25 nmoles |
| 17036 | Fluorescein-12-dATP *1 mM in Tris Buffer (pH 7.5)* | 25 nmoles |
| 17037 | Fluorescein-12-dGTP *1 mM in Tris Buffer (pH 7.5)* | 25 nmoles |
| 17038 | Cyanine 5-dATP [Cy5-dATP] | 25 nmoles |
| 17056 | Aminopropargyl dATP [7-Deaza-7-Propargylamino-2'-deoxyadenosine-5'-triphosphate] | 10 umoles |
| 17059 | Aminopropargyl dGTP [5-Propargylamino-2'-deoxyguanosine-5'-triphosphate] | 10 umoles |
| 17090 | 7-Deaza-7-Propargylamino-3'-azidomethyl-dATP | 50 nmoles |
Table 2. Deoxynucleotides (dNTPs) for use in PCR, real-time PCR, and reverse transcription PCR