Downstream manipulations of PCR products, such as restriction digests, ligation, sequencing or hybridization, often require purification of the amplified DNA. Alternatively, PCR products can be directly ligated in plasmids, transformed and propagated in bacteria. Use schematic illustration below to help plan your workflow.
Whether you choose our PCR purification tools and gel extraction kits to obtain DNA fragments with the right purity and concentration for the next step, or whether you opt for one of our easy subcloning kits to preserve your product, you can expect superior recovery, accuracy, and efficiency.
AccepTor™ Vector Kits
Easily and quickly clone PCR products without restriction digests or special primers. Use AccepTor vector kits to clone PCR products with single 3´-dA overhangs, which are generated by DNA polymerases such as KOD XL and Taq polymerases. The ready-to-ligate AccepTor Vector contains single 3´-dU ends. Simply mix the vector with your PCR product, incubate with Clonables™ 2X Ligation Premix, and transform into NovaBlue Singles™ Competent Cells—the entire process can be completed in just 40 minutes.
Perfectly Blunt Cloning Kits
Efficiently clone DNA with any type of end in less than an hour, with no restriction digests or special primers. Perfectly Blunt Cloning Kits are a convenient platform for subcloning DNA sequences amplified with KOD polymerase, which generates blunt ends. Alternatively, use the kit’s End Conversion Mix to to produce blunt, phosphorylated ends, which are compatible with the linearized, dephosphorylated blunt vector. This approach enables simple cloning after amplification with high-fidelity proof reading enzymes, which decreases the probability of mutations in the target sequence. In addition, blunt cloning can be more efficient than T-cloning. These kits are also perfect for cloning restriction fragments, cDNA, or sheared DNA.
Amicon® Ultra 0.5 mL Centrifugal Filters, MWCO 30,000
Centrifuging PCR reactions through an ultrafiltration membrane is a fast, easy way to separate DNA amplicons from other reaction components. With its engineered dead stop and vertical membrane design, the industry-leading Amicon Ultra filter provides the best yields with the fastest spin times, and enables simultaneous concentration and purification of DNA fragments.
Additional wash optimizes primer removal while maintaining yield when separating a PCR product from primers with Amicon Ultra 0.5 mL filters. 100 μL of PCR product in Tris-EDTA (TE) buffer was spiked with fluorescently labeled primers, and spun for 12 min. To half of the devices, an additional 500 μL of TE was added and spun again. After a final reverse spin, the amount of primers and PCR product (using fluorescence) was determined in each retentate and filtrate. The chart shows percent PCR product recovery and percent primer removal using either a single elution (left) or additional wash (right).
MultiScreen PCR Filter Plates
PCR filter plates are fast, automatable solutions for high-throughput PCR purification. Available in 96- and 384-well formats as well as micro 96-well format for small volumes, MultiScreen PCR plates provide high purity and recovery with a much shorter protocol and fewer handling steps than other methods. The plates use a size-exclusion membrane and vacuum filtration to provide a one-step protocol for excellent results. No centrifugation or precipitation steps are required.
Fast, high-throughput PCR clean-up in three
easy steps:
- Load PCR reactions.
- Filter with EMD Millipore vacuum manifold for 5–10 minutes or until wells are dry.
- Add water or buffer to each well. Agitate by shaking or pipetting. Retrieve purified samples by aspiration.
Proven workflow successes with below automated platforms:
- Biomek® FX Nucleic Acid Preparation System
- Packard MultiPROBE® II EX Liquid Handling System
- Beckman Multimek96 Pipettor
SpinPrep™ PCR Clean-Up Kit
Combine simplicity, speed, range and sensitivity when you use this kit to clean up and recover PCR-amplified DNA. The 3-step, 10-minute spin column procedure involves adsorption of DNA to silica membrane activated with binding buffer. After a wash step, DNA is eluted using a low-salt buffer. Effectively remove DNA polymerases, dNTPs, salts and primers from the PCR reaction mix to avoid interference with downstream applications such as cloning, sequencing or labeling. Purify PCR products from 100 bp to > 12 kbp with up to 90% recovery.
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| Column binding capacity: | up to 6 µg |
| PCR sample volume: | 100 µL/rxn |
| Typical recovery: | 60-90% |
| Size range: | 100 bp to > 12,000 bp |
| Time required: | < 10 min |
SpinPrep Gel DNA Kit
Quickly and efficiently extract DNA fragments from 150 bp to >12,000 bp from agarose gels without organic extraction or alcohol precipitation. The procedure uses GelMelt™ Solution to dissolve the gel slice, followed by adsorption of the DNA to a silica membrane in a spin column format. After a wash step, the purified DNA is eluted in low-salt buffer.
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| Column binding capacity: | up to 20 µg |
| Gel slice mass: | 150 mg/rxn |
| Typical recovery: | 50-90% |
| Size range: | 150 bp to > 12,000 bp |
| Time required: | < 30 min |
Gel analysis and quantification of DNA frag-ments isolated with the SpinPrep Gel DNA Kit
Known amounts (2 μg) of four DNA fragments of the indicated sizes were run in separate lanes on a 1% agarose gel. Each band was excised and the DNA extracted from the gel using the SpinPrep Gel DNA Kit and standard protocol. Recoveries shown as percentages above were determined by absorbance at 260 nm. Samples (250 ng) of each recovered band were analyzed by agarose gel electrophoresis. Lane 1 contained a mixture of the starting DNAs.
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