Necessity of Purified DNA and RNA

The uses of synthetic DNA and RNA are expanding across various fields, including clinical diagnostics and the development of novel biopharmaceutical therapeutics. DNA and RNA synthesis is a highly efficient process that generates minimal impurities at each step during the synthesis cycle. For example, it is crucial to purify RNAs obtained from enzymatic synthesis or modification reactions before utilization. After in vitro transcription, the RNA needs to be purified to remove unincorporated nucleotides, short aborted transcripts, enzymes, and buffer components. After performing protocols like RNA labeling, DNase I treatment, proteinase K treatment, and mRNA capping, it is crucial to eliminate small molecule and enzyme reaction components. This thorough removal to produce highly purified RNA is necessary for further applications or experiments, such as RNA vaccines.

Fig.1 Types of mRNA vaccines. (Esteban, et al., 2021)

Key Technologies

The key technologies of our DNA and RNA purification service include PAGE-based purification, which separates polynucleotides by size with high resolution, and HPLC-based purification employing three specialized methods: reversed-phase HPLC (separates by hydrophobicity), ion-exchange HPLC (separates by charge), and ion-paired reversed-phase HPLC (enhances resolution for charged molecules), ensuring high-purity outcomes suitable for sensitive downstream applications.

Purified Precision: Where DNA Meets RNA, Perfected by Glyco-Science.

This purified DNA and RNA can be used for diverse downstream applications. As a result, CD BioGlyco offers better methods to purify and analyze these vital biological aptamers to guarantee delivering DNA and RNA of high quality. We have two techniques available to purify and isolate DNA and RNA from synthetic products. These techniques are employed to eliminate reaction by-products and undesired oligonucleotides produced during the synthetic reactions. We primarily utilize PAGE and HPLC for the acquisition of desired compounds derived from synthetic products. We efficiently isolate the desired DNA and RNA and remove any unwanted by-products and impurities that may arise during the synthetic reactions through our techniques.

• PAGE-based DNA and RNA Purification Service
  We employ PAGE separation to purify DNA and RNA, which can be performed at either the analytical or preparative scale. PAGE technology separates polynucleotides based on their length, allowing for the differentiation of full-length molecules from truncated ones. Our technique of PAGE separation effectively separates and purifies the desired molecules while eliminating undesired impurities, ensuring a high level of purification for DNA and RNA samples.

Fig.2 The main factors determining the separation properties. (CD BioGlyco)

• HPLC-based DNA and RNA Purification Service
  HPLC, a reliable method for analyzing and purifying synthetic DNA and RNA, successfully achieves the separation of polymers with similar sizes and allows for automation.
  
  • Reversed-phase HPLC
    Reversed-phase HPLC separates polynucleotides and contaminants by exploiting variations in hydrophobicity. Crude polynucleotides consist of the target product and chemically modified by-products, all exhibiting different levels of hydrophobicity.
  • Ion-exchange HPLC
    Ion-Exchange HPLC is widely employed for the purification and separation of polynucleotides with multiple charges at CD BioGlyco. By utilizing a salt-gradient elution mode on a polymeric adsorbent or porous silica derivatized with tertiary or quaternary ammonium, the ion-exchange HPLC technique enables the effective separation of DNA and RNA.
  • Ion-paired reversed-phase HPLC
    Ion-paired reversed-phase HPLC is an analytical method that enhances resolution by incorporating a low concentration of long-chained alkyl amine into the mobile phase. This alkyl amine interacts with negatively charged oligonucleotides, thereby improving the separation of the analytes.

Workflow

Our streamlined purification workflow is designed for maximum efficiency, transparency, and quality assurance. Each project undergoes a meticulous, multi-stage process to ensure the highest possible nucleic acid yield and integrity.

• Sample Collection and Lysis

The process begins with the careful receipt and preparation of your samples. We employ a combination of physical, chemical, and enzymatic methods to disrupt cell membranes and release nucleic acids into a solution, creating a crude lysate.

• Lysate Clearing

The crude lysate is then processed to remove cellular debris, lipids, and proteins that could interfere with subsequent purification steps. This is a critical stage that ensures the integrity of the downstream purification process.

• Purification

Based on your project requirements, we apply either our PAGE or HPLC purification protocols. For PAGE, we first visualize the DNA or RNA band of interest, carefully excise it from the gel, and then recover the nucleic acid from the gel matrix. For HPLC, the sample is injected into a column where it binds to a resin and is then eluted in a stepwise manner to separate the target molecules.

• Washing and Elution

After purification, the target nucleic acids are washed to remove residual salts, buffers, and other contaminants. The final, highly purified DNA or RNA is then eluted in a low-ionic-strength solution, such as nuclease-free water, to ensure its readiness for your specific downstream application.

• Quality Control and Reporting

Every purified sample undergoes rigorous quality control. We provide comprehensive reports detailing the concentration, purity, and integrity of your nucleic acids, giving you a complete overview of the quality of your sample.

Publication Data

Applications

• PAGE can be applied in preparative scenarios, including the small-scale purification of radioactive single-stranded probes and the large-scale purification of synthetic polynucleotides.
• HPLC offers exceptional purification capabilities for a wide range of compounds, including (5'-monomethoxytrityl 1, 5'-dimethoxytrityl 1,2, fluorescein, phosphorothioate 4, tert-butyldimethylsilyl 5,7, and N-acetyl 2-aminofluorene 3,10) protected and deprotected oligomers. These columns ensure excellent recovery rates, allowing for efficient isolation and purification of these compounds while maintaining their structural integrity.

Advantages

• The application of PAGE for the separation of polynucleotides larger than 50-60 m in length is well-established and highly efficient. This technique provides exceptional resolution and allows for precise separation of these extended nucleotide sequences. It is important to understand their structures and functions.
• HPLC is directly compatible with MS instruments and that is easy for the identification of individual peaks.
• This coupling provides for the efficient characterization and analysis of compounds within the sample. That provides valuable information on their molecular structures, composition, and fragmentation patterns.

Frequently Asked Questions

Associated Services

Our DNA and RNA purification delivers high-integrity nucleic acids essential for genomics and transcriptomics studies. To unlock deeper insights into glycan-mediated biological recognition, critical for understanding infection, immunity, and cellular communication, we provide advanced Glycan Microarray Assays. These platforms rapidly profile interactions between carbohydrates and diverse biomolecules (e.g., antibodies, lectins, viruses), enabling high-throughput functional glycomics:

• Synthetic Glycan Microarray Assay
• Sialylated Oligosaccharides Microarray Assay
• Glycosaminoglycan (GAG) Microarray Assay
• ABH Blood Type I-VI Antigen Microarray Assay

As a leading company in the field of glycobiology, CD BioGlyco excels in DNA and RNA purification. We possess several advantages that set us apart from others. We employ state-of-the-art purification techniques to ensure efficient and pure extraction of the target molecules. Besides, our research team possesses extensive knowledge and experience in the field. If you would like to know more about our services, we encourage you to contact us for further information.

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