Nanosep Centrifugal Ultrafiltration Devices: Uses for Cloning and PCR

28 Aug 2017

Molecular cloning is a technique often used in most areas of biological research. Standard molecular cloning techniques require enzymatic modification and purification steps to prepare DNA sequences for litigation and transformation into bacterial cells. The polymerase chain reaction (PCR) is employed to generate more of the desired DNA fragment ready for cloning. The use of Nanaosep® centrifugal ultrafiltration devices in various steps of the cloning process can provide a more streamlined process compared to more traditional methods. This application note demonstrates how DNA products from PCR were concentrated and desalted while simultaneously allowing primers and adapter fragments to pass into the discarded filtrate, saving time and materials while providing results similar to those obtained via traditional methods.

Nanosep® & Nanosep MF Centrifugal Devices

Pall Life Sciences - Laboratory, Food, Beverage

Simple, reliable concentrating and desalting of 50 to 500 µL samples. Ensures rapid processing of samples. Typical recoveries are greater than 90%. Available with low protein binding Omega™, Bio-Inert®, and GHP membranes. A wide range of MWCOs, color-coded for easy identification. Constructed of low-binding polypropylene. Ultrasonically welded seals prevent bypass or seal failure. Fits standard centrifuge rotors that accept 1.5 mL tubes.

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PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.MicrobiologyMicrobiology is the study of microorganisms including protists, prokaryotes, fungi, and, often, viruses. Microorganisms are a useful research tool as genetic vectors and, in immunology, for antibiotic susceptibility testing, cellular biology and genetics. Microorganisms commonly grow readily in incubators with microbial culture media; this can contain chromogenic supplements to differentiate between cell lines. Estimate your culture’s density of microorganisms with colony counters, or screen and select colonies for desirable clones with automated colony pickers. Additionally, equipment is available to monitor environments for the presence of microbes and identify with microbial identification instruments. Find the best microbiology products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Nucleic Acid ConcentrationUltrafiltration