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The Crucial Role of dNTPs in cDNA Synthesis

Explore the pivotal role of dNTPs in cDNA synthesis, unraveling the intricate process where these molecular building blocks lay the foundation for converting RNA into complementary DNA strands

July 13, 2023

In the realm of molecular biology, the synthesis of complementary DNA (cDNA) stands as a fundamental technique with widespread applications, from gene expression analysis to understanding the intricacies of cellular processes. At the heart of cDNA synthesis lies a crucial player: deoxynucleoside triphosphates (dNTPs). This article delves into the significance of dNTPs in the intricate process of cDNA synthesis.

Understanding cDNA Synthesis

cDNA synthesis is a pivotal technique that involves the conversion of RNA molecules into DNA. It serves as a cornerstone in various molecular biology applications, facilitating the study of gene expression and regulatory mechanisms. The process entails the utilization of a reverse transcriptase enzyme, which catalyzes the synthesis of a complementary DNA strand using an RNA template.

The Role of dNTPs

Central to the process of cDNA synthesis are dNTPs – the building blocks of DNA. Each dNTP molecule encompasses one of the four nitrogenous bases – adenine, guanine, cytosine, or thymine – along with a deoxyribose sugar and three phosphate groups. These molecules serve as the substrates for the reverse transcriptase enzyme during cDNA synthesis.

Incorporation and Complementary Base Pairing

During cDNA synthesis, dNTPs are incorporated into the growing DNA strand through complementary base pairing with the template RNA. For instance, if the template RNA harbors an adenine base, the reverse transcriptase enzyme will add a thymine-containing dNTP to the growing DNA strand. This process continues until the entire RNA template is transcribed into a complementary DNA strand.

Ensuring Efficiency and Accuracy

The presence of dNTPs is indispensable for efficient and accurate cDNA synthesis. Proper concentration and balance of dNTPs are vital to ensure optimal DNA synthesis and to prevent errors or biases in the resulting cDNA library. Typically, dNTPs are provided in excess during cDNA synthesis reactions to ensure an ample supply of building blocks for DNA synthesis.

Conclusion

In essence, dNTPs serve as the backbone of cDNA synthesis, facilitating the conversion of RNA into complementary DNA strands. Their precise incorporation through complementary base pairing ensures the fidelity and accuracy of the synthesized DNA. As researchers continue to explore the depths of molecular biology, the role of dNTPs in cDNA synthesis remains paramount, driving advancements in various fields ranging from genetics to medicine.

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).