PAGE-based DNA and RNA Purification Service
Overview of PAGE
DNA and RNA are a class of macromolecules with important functions for living organisms, and there is often a requirement for high purity for their use. Therefore, it is a very urgent matter to comprehensively study the way of separating and purifying biomolecules. Among the techniques for the separation and purification of biological macromolecules, gel electrophoresis is a basic technique in the laboratory of biological disciplines. This technique allows efficient separation of macromolecules such as DNA, RNA, and proteins. For proteins in particular, polyacrylamide gel electrophoresis (PAGE) is often the technique of choice. The separation principle of PAGE is based on the electrophoretic mobility of biomolecules (the ability of an analyte to move toward an electrode of opposite charge). In the use of PAGE, the charge, size (molecular weight), and shape of the molecule determine the specific operation.
Key Technologies
The key technology of this service is PAGE, which utilizes an electric field to separate DNA and RNA fragments based on their electrophoretic mobility, influenced by size, charge, and shape, under denaturing conditions for high-resolution purification and subsequent extraction of target molecules from the gel.
PAGE-Perfected: Where Every Base Meets Its Band.
For the use of PAGE purification technology, CD BioGlyco has many years of operational experience. The PAGE purification method we offer mainly utilizes denaturing polyacrylamide gel electrophoresis for the separation of primed DNA / RNA, followed by the recovery of the target DNA / RNA from the gel. Currently, the PAGE purification method we offer provides greater than 90% purity of purified DNA and is particularly effective for the purification of long-stranded DNA (greater than 50 mer).
The PAGE purification technique is suitable for the isolation and purification of large polynucleotide DNA or RNA fragments and allows the isolation of relatively small proteins.
- Specifically, CD BioGlyco offers PAGE purification techniques that apply to situations:
- Primers for routine molecular biology experiments
- Long-chain macromolecules (≥60 mer)
- Unmodified oligonucleotides over 50 mer
- Most long strands of DNA or RNA
- Depending on the purpose of the analysis, we offer PAGE including:
- SDS PAGE
- Native PAGE
- Buffer used
- SDS PAGE uses trimethylene glycol, bis-trimethylene glycol, triacetate, and tris-tri ethylene glycol, of which trimethylene glycol is the most commonly used.
- Native PAGE uses TBE without SDS.
Fig.1 Flow chart of PAGE purification technology. (CD BioGlyco)
Workflow
Publication Data
DOI.: 10.1021/acs.oprd.4c00382
Journal: Organic Process Research & Development
IF: 3.5
Published: 2024
Results: This review details PAGE as a traditional method for purifying synthetic oligonucleotides based on length/size separation. PAGE effectively isolates full-length products from failure sequences (e.g., n–1 impurities) by exploiting differential migration through a gel matrix under an electric field. While advantageous for analytical and low-preparative scales (≤1 mmol), PAGE exhibits critical limitations: low recovery rates (<50%) due to post-electrophoresis extraction and desalting steps, inability to resolve chemically modified nucleosides (e.g., 2′-OMe, 2′-F), and poor scalability. The method is also labor-intensive and time-consuming, requiring extensive manual handling. Despite its historical utility for research-grade oligonucleotides, PAGE is largely superseded by chromatographic techniques (e.g., ion-exchange, reversed-phase) for therapeutic applications demanding higher purity, yield, and scalability.
Applications
- PAGE could be used for the purification and analysis of DNA and RNA.
- PAGE could be used for quantitative and structural analysis of proteins.
- PAGE could be used to identify nucleic acid-protein complexes.
Advantages
- Our PAGE technology delivers superior resolution, effectively separating full-length nucleic acids from synthesis byproducts, truncated fragments, and other contaminants.
- Our meticulous workflow and extensive experience ensure that each sample is processed with the highest level of precision, providing reliable and consistent results batch after batch.
- Our team of specialists provides personalized consultation and support, guiding you through the selection of the best purification method for your specific project.
Frequently Asked Questions
Associated Services
Our PAGE-based DNA and RNA purification offers high-resolution size selection for nucleic acid fragments, ideal for applications demanding precise molecular weight isolation—from CRISPR guide RNAs to synthetic biology constructs. To transform these purified sequences into functionally annotated tools for decoding glycan-protein interactions, we offer LiGA Construction Services. This platform genetically engineers multivalent glycan probes for live-cell or in vivo binding studies, enabling:
CD BioGlyco has many years of experimental experience in the purification of DNA and RNA. We have professional researchers and first-class experimental equipment to provide custom isolation and purification solutions for clients all over the world. At the same time, we also provide timely feedback to our clients on the progress of their experiments. If you are interested in our services, please feel free to contact us.
Reference
- Abe, A.; Časar, Z. Overview and recent advances in the purification and isolation of therapeutic oligonucleotides. Organic Process Research & Development. 2024, 29(1): 15-33. (Open Access)
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