Polymerase Chain Reaction (PCR) has revolutionized molecular biology and various fields reliant on genetic analysis. At the heart of PCR lies the strategic interplay of several crucial components, including primers, DNA polymerase, template DNA, and dNTPs (deoxyribonucleotide triphosphates). Among these, dNTPs serve as the building blocks for synthesizing new DNA strands during PCR.
The concentration of dNTPs in a PCR reaction profoundly influences the efficiency and fidelity of DNA amplification. While there isn't a one-size-fits-all concentration, optimal dNTP levels typically range between 0.2 to 0.4 mM (millimolar). This range ensures sufficient substrate availability for DNA polymerase while minimizing non-specific amplification and primer-dimer formation.
Achieving the ideal dNTP concentration involves careful consideration of various factors. Firstly, the choice of DNA polymerase dictates the permissible dNTP concentration range. High-fidelity polymerases may require lower dNTP concentrations to reduce the risk of misincorporations, while less proofreading polymerases might benefit from higher dNTP levels to compensate for their error-prone nature.
Additionally, the length and complexity of the target DNA sequence influence dNTP optimization. Longer amplicons or regions with high GC content may necessitate higher dNTP concentrations to ensure efficient amplification. Conversely, shorter targets or sequences prone to secondary structure formation may perform optimally with lower dNTP concentrations to mitigate nonspecific amplification.
Furthermore, the presence of potential inhibitors in the reaction mixture warrants adjustments to dNTP concentrations. Suboptimal dNTP levels can impede PCR efficiency, leading to failed or subpar amplification. Therefore, it's crucial to assess the compatibility of dNTP concentrations with reaction components and optimize accordingly.
Optimizing dNTP concentration often involves a systematic approach of testing different concentrations within the established range. Researchers may employ gradient PCR or serial dilution methods to evaluate the impact of varying dNTP concentrations on amplification efficiency, specificity, and yield. By systematically assessing these parameters, researchers can identify the optimal dNTP concentration for their specific PCR assay.
Moreover, advancements in PCR technology and the availability of specialized dNTP formulations offer additional avenues for optimization. Modified nucleotides, such as dUTP or fluorescently labeled dNTPs, cater to specific applications like quantitative PCR (qPCR) or genotyping assays, allowing for enhanced sensitivity and multiplexing capabilities.
In conclusion, optimizing dNTP concentration in PCR reactions is a critical step in ensuring robust and reliable DNA amplification. By considering factors such as polymerase fidelity, target sequence characteristics, and potential inhibitors, researchers can tailor dNTP concentrations to maximize PCR performance. Through systematic optimization strategies and leveraging technological advancements, PCR assays can achieve heightened efficiency, specificity, and versatility, advancing applications in research, diagnostics, and beyond.
Why Choose SBS Genetech dNTPs for PCR?
At SBS Genetech, we recognize the pivotal role that dNTPs play in molecular biology experiments, particularly in PCR reactions, where they serve as essential keystones. The efficiency and accuracy of PCR amplification hinge directly upon the quality and concentration of dNTPs. It's with this understanding that we've meticulously crafted our molecular biology grade dNTPs, ensuring they meet the highest standards of quality and reliability.
Quality Assurance Beyond Measure
Our commitment to excellence drives us to subject our dNTPs to rigorous testing and verification processes. Through meticulous quality control measures, we guarantee that our dNTPs exhibit purity levels of up to 99% as assessed by High-Performance Liquid Chromatography (HPLC). This purity, coupled with the absence of contaminants such as RNase and DNase, makes our dNTPs ideal for a myriad of molecular biology applications.
Trusted by Researchers, Cited in Literature
The efficacy of our dNTPs is not merely a claim but a testament backed by the trust of countless researchers worldwide. Widely employed in diverse molecular biology research endeavors, our dNTPs have become indispensable tools for scientists striving for precision and reproducibility in their experiments. Moreover, their reliability is underscored by their frequent citation in related literature, further solidifying their reputation as a gold standard in the field.
Features Designed for Excellence
Our dNTPs boast features designed to empower researchers and elevate their work:
- Ultra-Pure Composition: With purity levels exceeding 99% as verified by HPLC, our dNTPs ensure the integrity of molecular biology experiments, delivering reliable and consistent results.
- Available in Convenient Formats: Whether as a ready-to-use mix or individual components, our dNTPs offer flexibility to suit diverse experimental needs, providing convenience without compromising quality.
Versatile Applications, Uncompromising Performance
The versatility of our dNTPs extends across a spectrum of molecular biology applications, including but not limited to:
- PCR and qPCR
- cDNA synthesis
- Primer extension
- DNA sequencing
- DNA labeling
- Mutagenesis
Storage Recommendations for Longevity
To preserve the integrity and stability of our dNTPs, proper storage conditions are imperative. We recommend storing them at -70°C for infrequent use or -20°C for daily or weekly use, guarding against freeze-thaw cycles that could compromise product stability.
Empowering Your Research Journey
At SBS Genetech, we're not just providers of dNTPs; we're partners in your research journey. Committed to facilitating scientific advancement, we strive to equip researchers with the highest quality tools to unlock new discoveries and drive innovation. Trust in SBS Genetech dNTPs to power your pursuit of scientific excellence.
Explore Our Comprehensive Product Range
Beyond dNTPs, we offer a comprehensive range of Ribonucleotides and Deoxynucleotides to cater to diverse research needs. For inquiries or to place an order, please reach out to tech@sbsbio.com.
In conclusion, SBS Genetech dNTPs stand as a beacon of quality, reliability, and precision in molecular biology research, empowering scientists to push the boundaries of knowledge and pave the way for groundbreaking discoveries.
Featured Citations
Interested in seeing published research using our dNTPs?
Visualized RNA detection of SARS-CoV-2 in a closed tube by coupling RT-PCR with nested invasive reaction
Analyst | 4 Jan 2023 | DOI: https://doi.org/10.1039/d2an01679f
The 20 μL reaction mixtures of the assay contained 1× visualized closed-tube PCR buffer (10 mM Tris–HCl (pH 8.5), 7.5 mM MgCl2·6H2O, 30 mM NaCl, 0.05% NP-40, 0.05% Tween-20), 50 U HiScript II reverse transcriptase, 0.25 mM dNTPs (SBS Genetech Co. Ltd, Beijing, China), 0.5 μM forward primer, 0.5 μM reverse primer, 0.25 U GoTaq DNA polymerase (Promega, Beijing, China), 3.5% PEG8000 (BSK Technology Co. Ltd, Nanjing, China), 0.1 μM UP, 0.4 μM DP, 0.2 μM hairpin probe, 100 ng of FEN1 endonuclease (prepared in our laboratory.
CRISPR/Cas genome editing perspectives for barley breeding
Psysiologia Plantarum | 22 Apr 2022 | DOI: https://doi.org/10.1111/ppl.13686
Primers for sgRNA of eIF4E genes were selected with WhU6 promoter region for amplification of a 362-base pair fragment: F 5′-GACCAAGCCCGTTATTCTGAC-3′, R 5′-AAGTCTGATGCAGCAAGCGAG-3′; for the region including Cas9 with the promoter: F 5′-GCTCCTGGTCCATCCACG-3′, R 5′-CGTG-GATGGACCAGGAGC-3′; for hptII: F 5′-GCTGCGCCGATGGTTTCTACA-3′, R 5′-GCCCAAAGCATCAGCTCATCG. The recommended amplification mixture contained 5 mg of the DNA template (Applied Biosystems); 2.5 mM MgCl2; 250 μM dNTPs (Beijing SBS Genetech Co., Ltd.)
Femtomolar and locus-specific detection of N6-methyladenine in DNA by integrating double-hindered replication and nucleic acid-functionalized MB@Zr-MOF
Journal of Nanobiotechnology | 7 Dec 2021 | DOI: https://doi.org/10.1186/s12951-021-01156-0
Klenow Fragment DNA polymerase (3′ → 5′ exo−), 10 × Klenow buffer (500 mM Tris–HCl, 50 mM MgCl2 and 10 mM DTT, pH 7.9), and 10 × CutSmart™ buffer (20 mM Tris–acetate, 500 mM potassium acetate, 10 mM magnesium acetate and 100 µg/mL BSA, pH 7.9) were obtained from New England Biolabs (Beijing, China). GoldView I, 20 bp DNA marker, and dATPs, dTTPs, dCTPs and dGTPs were purchased from SBS Genetech Co., Ltd., (Beijing, China)
Multiplex Visualized Closed-Tube PCR with Hamming Distance 2 Code for 15 HPV Subtype Typing
Anal. Chem. | 22 Mar 2021 | DOI: https://doi.org/10.1021/acs.analchem.1c00035
Reagents included GoTaq Hot Start Polymerase (Taq DNA polymerase) (Promega), flap endonuclease 1 (FEN1) prepared in our laboratory as described previously, (19) deoxynucleotide triphosphates (dNTPs) (SBS Genetech Co., Ltd., China)
An integrated electrochemical biosensor based on target-triggered strand displacement amplification and “four-way” DNA junction towards ultrasensitive detection of PIK3CA gene mutation
Biosensors and Bioelectronics | 15 Feb 2020 | DOI: https://doi.org/10.1016/j.bios.2019.111954
NsbI restriction enzyme, Klenow Fragment (KF) (3′→5′exo-), Nb.BbvCI, 10 × Klenow buffer (500 mM Tris-HCl, 50 mM MgCl2 and 10 mM DTT, pH 7.9) and 10 × CutSmart™ buffer (20 mM Tris-acetate, 500 mM potassium acetate, 10 mM magnesium acetate and 100 μg/mL BSA, pH 7.9) were obtained from New England Biolabs (Beijing, China). GoldViewⅠ, DNA marker and dNTP were purchased from SBS Genetech Co., Ltd (Beijing, China)
Sequence-encoded quantitative invader assay enables highly sensitive hepatitis B virus DNA quantification in a single tube without the use of a calibration curve
Royal Society of Chemistry | 8 Aug 2019 | DOI: https://doi.org/10.1039/c9an00970a
A virus RNA/DNA Extraction Kit was purchased from Xi'an Tianlong Science and Technology Co., Ltd (Xi'an, China), deoxynucleotide triphosphates (dNTPs) were obtained from SBS Genetech Co., Ltd (Beijing, China)
Dual cycle amplification and dual signal enhancement assisted sensitive SERS assay of MicroRNA
Analytical Biochemistry | 1 Jan 2019 | DOI: https://doi.org/10.1016/j.ab.2018.10.004
Klenow fragment of E.coli DNA polymerase and nicking endonuclease (NEase) were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). BEAS-2B cells was purchased from GeFan Biotechnology.Go.,Ltd (Shanghai, China). Cell lysis buffer was purchased from Sangon Biotech (Shanghai, China). The mixture of four dNTPs (10 mM for each component) was purchased from SBS Genetech Co., Ltd. (Beijing, China).