Workflow Improvement for PCR-based Genetic Testing: Protocol Consolidation, Decreased Cycle Times, and Increased Pass Rates

14 Aug 2014

This case study documents a project to consolidate PCR protocols and reduce costs, using a KAPA workflow at a large genetic testing facility. The study shows that the KAPA2G Robust HS ReadyMix formulation has potential to streamline workflow, increase pass rates, and simplify reaction setup.

KAPA Express Extract Kits

Kapa Biosystems, Inc.

Rapid and efficient extraction of DNA from a broad range of source material. KAPA Express Extract is a novel thermostable protease and buffer system that allows for the extraction of PCR-ready DNA from various tissue types in as little as 15 minutes. The KAPA Express Extract system has been designed for optimal tissue lysis and DNA preservation. DNA extractions are conveniently performed in a single-tube, without the need for hazardous chemicals and multiple washing steps, thereby greatly reducing the risk of sample loss and contamination. Key features of the KAPA Express Extract system include: Rapid extraction protocol - PCR-ready DNA in 15 minutes Versatility - single kit optimized for a variety of sample types Single-tube reaction minimizes risk of contamination Couple with KAPA2G Robust HotStart ReadyMix for increased PCR success rates The combination of KAPA Express Extract and KAPA2G Robust HotStart ReadyMix provides the highest performance solution for rapid DNA extraction and consistent downstream amplification. KAPA2G Robust DNA Polymerase is a novel enzyme evolved for increased processivity and exhibits greatly improved tolerance to carryover PCR inhibitors from crude DNA extractions.

(1)

KAPA2G Robust PCR Kits

Kapa Biosystems, Inc.

Streamline your PCR workflows by consolidating protocols and reaction conditions, increase pass rates, amplify difficult templates without bias, and increase sensitivity – all with a single enzyme. The second-generation KAPA2G Robust DNA Polymerase was evolved to solve - achieve consistent amplification across a broad range of amplicon types (both GC and AT-rich). The versatility and robustness of the polymerase allows for consolidation of PCR cycling protocols and reaction conditions while increasing success rates. The high performance of the KAPA2G Robust DNA Polymerase eliminates the need for multiple enzymes and protocols - standardize your PCRs with a single enzyme solution. The KAPA2G Robust DNA Polymerase offers: Robust performance across a wide range of GC- and AT-rich templates Increased PCR success rates Consolidated PCR protocols and reaction conditions with a single enzyme Improved tolerance to common PCR inhibitors for crude samples and/or DNA extractions Consistent Colony PCR from E. coli and yeast Higher yield and sensitivity per unit of enzyme Three novel buffers and a proprietary PCR enhancer offer extended optimization options for the most difficult templates KAPA2G Robust DNA Polymerase is also available with HotStart technology for improved specificity during prolonged benchtop setup or liquid handling.

(1)

Links

Tags

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.HematologyIn Haematology / Hematology, complete blood cell counts (or full blood counts) are obtained using automated blood count analyzers to enumerate blood cell types.  Hematology also encompasses haemostasis and coagulation, thrombophilia and hemophilia, plasma viscosity and ESR analysis, hemoglobinopathies, cell morphology and haematinic measurement.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.DNA SequencingDNA sequencing, such as sanger sequencing, is a biological technique that determines the precise order of nucleotide bases in a fragment or template of DNA. DNA sequencers and genetic analyzers are based on capillary electrophoresis, where labeled DNA fragments are electrophoretically separated by size as they migrate through a polymer. Find the best DNA sequencing products, including DNA sequencing kits, genomic libraries and genetic identity kits in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Clinical GeneticsMolecular Genetics covers the analysis of hereditary genetic disease and chromosomal abnormalities. Genetics can be analysed using DNA, RNA, and protein microarrays, PCR, RT PCR and DNA sequencing. Genetic equipment includes genetic workstations, thermal cyclers, cooling blocks and electrophoresis products. Diagnostic kits are used for DNA / RNA extraction and purification.Next Generation SequencingNext-generation sequencing (NGS), also known as whole-genome sequencing, high-throughput sequencing and massive parallel sequencing, produces and analyses thousands to millions of nucleotide sequences at once. Sequencing systems operate via varying technologies depending on the manufacturer, including sequencing by synthesis, ligation, pyrosequencing, ion semiconductor and single-molecule real-time sequencing. For NGS, library preparation is paramount to successful sequencing. In this section, explore a range of library preparation kits, from targeted, amplicon-based or hybridization-based kits including epigenomic, transcriptomic and genomic workflows to fragmentation kits. Find the best next-generation sequencing products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.PCR workflow