Deoxynucleotide triphosphates (dNTPs) and cyclic-di-AMP Analysis Service

The importance of dNTPs

The dNTPs, or deoxynucleotide triphosphates, including dATP, dGTP, dCTP, dTTP and dUTP, are vital for various biological processes. The dATP, dGTP, dCTP, and dTTP serve as the fundamental constituents for DNA synthesis, furnishing the essential precursors for DNA replication and repair. During DNA replication, dNTPs are incorporated into the growing DNA strand by DNA polymerase enzymes, ensuring accurate and faithful duplication of the genetic material. Worth noting, the presence of dNTPs existing in cells are crucial in maintaining the correct levels for minimizing the mutation rate and preventing genome instability. However, an imbalance, either deficiency or surplus, of a single dNTP can lead to increased mutation rates during DNA replication. Furthermore, dNTPs play a crucial role in cellular metabolism and energy transfer. They are involved in ATP synthesis and serve as energy carriers in various biochemical reactions. Additionally, dNTPs are essential for DNA repair mechanisms, enabling the correction of errors and damage in the DNA sequence. Overall, the importance of dNTPs lies in their fundamental role in DNA synthesis, replication, repair, and cellular metabolism, making them essential components for the maintenance of genetic integrity and cellular functions, as improper or imbalanced dNTPs pools may lead to growth defects and oncogenesis.

Figure 1. Coupled synthesis of dNTPs and DNA ^[1].

The importance of cyclic-di-AMP ^[2]

Cyclic-di-AMP is an important signaling small molecule in various bacteria. Its significance lies in its role in regulating essential cellular processes, including osmotic stress response and cell wall homeostasis, virulence, and antibiotic resistance. In bacteria, this molecule helps maintain cellular integrity and osmotic balance, especially under fluctuating environmental conditions, making it crucial for bacterial survival and pathogenesis. Moreover, cyclic-di-AMP has been linked to bacterial virulence, as it influences the ability of certain pathogens to invade host cells and cause infections. Its regulatory role in virulence factors underscores its significance as a potential target for antimicrobial strategies. Furthermore, cyclic-di-AMP contributes to antibiotic resistance in some bacterial species. Its involvement in regulating cell wall synthesis can impact the efficacy of antibiotics targeting the bacterial cell envelope, making it a relevant factor in the context of antimicrobial resistance mechanisms. In summary, it is now recognized as a near-ubiquitous second messenger that coordinates multifaceted role of bacterial growth and behavior, including motility, virulence, biofilm formation and cell cycle progression. As a signaling molecule that influences key bacterial processes, including osmotic regulation, cell wall homeostasis, virulence, and antibiotic resistance. Understanding its functions is crucial for formulating strategies to address bacterial infections and combat antimicrobial resistance.

Figure 2. Cyclic nucleotide metabolism [3].

Our deoxynucleotide triphosphates (dNTPs) and cyclic-di-AMP Analysis Service

Up to now, various methods have been developed for quantifying cellular levels of dNTPs, including enzymatic and liquid chromatography assays. The latter involves direct separation of dNTPs by high-performance liquid chromatography (HPLC), which can be detected using UV or tandem mass spectrometry. Ideally, a combination of these two methods should be used for sensitive and reliable quantification. Similarly, the identification and quantification of cyclic-di-AMP would be more effectively achieved through the utilization of LC-MS/MS. With integrated set of separation, characterization, identification, and quantification systems featured with excellent robustness & reproducibility, high sensitivity, Creative Proteomics provides reliable, rapid, and cost-effective deoxynucleotide triphosphates (dNTPs) and cyclic-di-AMP targeted metabolomics services.

Technological superiority

1. Professional detection and analysis capability: Experienced research team, strict quality control system, together with ultra-high resolution detection system and professional data pre-processing and analysis capability, ensure reliable and accurate data.

2. Reproducible: Obtain consistent and reproducible inter- and intra- assay results for data analysis.

3. High veracity of data: For sample dNTPs and cyclic-di-AMP identification, qualitative and quantitative analysis can be achieved efficiently and accurately, with cost-effective, and standard curve R²>0.99.

4. High resolution and sensitivity: AB SCIEX QTRAP 6500 Plus, AB SCIEX QTRAP 5500, et al.

Samples Requirement

• Normal Volume: 200 μL serum/plasma; 200 mg tissue, 2×10⁷ cells
• Minimal Volume: 50 μL serum/plasma; 50 mg tissue, 5×10⁶ cells.
• Any other samples such as body fluid, feces, cell culture medium supernatant.

Results Delivery

1. A detailed technical report will be provided at the end of the whole project, including the experiment procedure, MS/MS instrument parameters, etc.

2. Raw data and data analysis results.

• Analytes are reported as μM or μg/mg (tissue), and variable-coefficient are generally<10%
• The name of the analytes, abbreviation, formula, molecular weight, and CAS# would also be included in the report.

How to place an order

At Creative Proteomics, many excellent and experienced experts will optimize the experimental protocol according to your requirement and guarantee the high-quality results for deoxynucleotide triphosphates (dNTPs) and cyclic-di-AMP Analysis Service. Please feel free to contact us by email to discuss your specific needs. Our customer service representatives are available 24 hours a day, from Monday to Sunday.

References

1. Loan TD, Easton CJ, Alissandratos A. DNA amplification with in situ nucleoside to dNTP synthesis, using a single recombinant cell lysate of E. coli. Sci Rep. 2019 Oct 30;9(1):15621.
2. Turner MS, Xiang Y, Liang ZX, et al. Cyclic-di-AMP signalling in lactic acid bacteria. FEMS Microbiol Rev. 2023 May 19;47(3): fuad025.
3. Rehmann H, Wittinghofer A, Bos JL. Capturing cyclic nucleotides in action: snapshots from crystallographic studies. Nat Rev Mol Cell Biol. 2007 Jan;8(1):63-73.