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DNA/RNA Clean-Up and Concentration

PCR products and DNA from restriction digests are commonly purified away from reaction components to remove any nucleotides or and primers or small molecules that can interfere with performance in downstream applications. The products listed here provide flexible options to purify and concentrate DNA or RNA fragments from PCR and from gels, or to concentrate DNA or RNA from dilute solutions.

The Wizard® SV Gel and PCR Clean-Up System extracts and purifies DNA fragments from agarose gels, or directly from PCR amplification reactions. ReliaPrep™ RNA and DNA Clean-Up and Concentration Systems can be used to concentrate dilute samples in a simple, fast protocol, or to purify DNA or RNA from amplification reactions and gels. The purified/concentrated DNA can be used for sequencing, cloning, labeling, restriction enzyme digestion or in vitro transcription/translation without further manipulation.

The Wizard® MagneSil® Sequencing Reaction Clean-up System provides a convenient magnetic-based method for purification of sequencing reactions that can be easily scaled for high-throughput applications.

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Introduction to DNA/RNA Clean-Up and Concentration

Purification or concentration of DNA fragments or PCR products involves separation of DNA from in vitro reaction components or from agarose gel slices. In many cases, after a PCR amplification or restriction enzyme digestion, the reaction components include primers, proteins and salts that may inhibit performance in subsequent analyses. 

Fragment DNA clean-up can improve efficiency in downstream applications. For example,  non-specific PCR products and primer-dimers in an amplification reaction can compete for ligation with the desired PCR product in subsequent T-vector cloning, resulting in low frequency of positive clones.

In Sanger sequencing, certain reaction components will interfere with data collection if not removed before data analysis. Buffer salts give rise to aberrant electrophoretic migration, and unincorporated dye-labeled terminators and related species interfere with the legitimate signal from sequencing extension products. 

Like other DNA and RNA purification systems, products designed to purify fragments or concentrate DNA or RNA typically involve immobilization of the nucleic acid on silica or cellulose incorporated into membranes, resins or magnetic beads, followed by washing and elution steps.