Using SapphireAmp® Fast PCR Master Mix for Rapid Colony PCR Screening

24 Jul 2014

This application note demonstrates the successful use of SapphireAmp Fast PCR Master Mix to identify plasmids containing a cloned fragment of the mouse CYP27A1 gene using a fast colony PCR screening strategy. PCR was completed in less than 60 minutes and amplification products detected to confirm the presence of the CYP27A1 insert.

SapphireAmp® Fast PCR Master Mix

Takara Bio

SapphireAmp Fast PCR Master Mix is a hot start PCR master mix that contains a hot start PCR enzyme, optimized buffer, dNTP mixture, gel loading dye (blue), and a density reagent in a convenient 2X premix. SapphireAmp PCR Master Mix is especially designed for fast PCR reactions that can be completed in half the time required when using conventional Taq DNA polymerase. For human genomic DNA, amplification of 2 kb targets can be completed in 1 hour, and amplifications of up to 6 kb are possible.This hot start PCR master mix product is especially well-suited for E. coli-based colony PCR to screen inserts up to ~5 kb in length. Colony PCR with SapphireAmp Master Mix can be completed in about 1 hour.SapphireAmp reactions can be assembled by simply adding primers and DNA template to the hot start PCR master mix. Reactions can be analyzed directly without purification; PCR products can be loaded directly onto a gel for electrophoresis or digested with restriction enzymes in the SapphireAmp reaction buffer.Features: Hot start PCR master mix is optimized for fast PCR Colony PCR can be completed in half the time required for conventional Taq DNA polymerase Convenient- eliminates purification steps with no reduction in yield Restriction enzyme digestion of the PCR products can be performed directly in the PCR buffer Reactions can be directly loaded onto a gel without purification or the addition of other reagents Applications: Hot start PCR Colony PCR Fast PCR

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Genome AnalysisGenomics, the study of genomes, includes functional genomics, evolutionary genomics and comparative genomics. There are many genomic technologies such as DNA sequencing of whole genomes, computational biology and bioinformatics. DNA and nucleic acids must be isolated and concentrated from cells for analysis with kits, automated analyzers and software. Other useful technologies for studying genomics include PCR, microarrays and electrophoresis.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.Molecular BiologyMolecular biology is the branch of biology that focuses on the molecular mechanisms that underlie cellular functions. It involves studying DNA, RNA, and proteins to understand gene expression, replication, and regulation. Molecular biology is fundamental to biotechnology, medicine, and genetic research. Explore molecular biology products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.ScreeningUsing robotics, data processing and control software, liquid handling devices and sensitive detectors, screening allows a researcher to quickly conduct millions of chemical, genetic or pharmacological tests.