Direct PCR on Hair: A New Animal-Friendly Genotyping Method

5 Apr 2016

Gene transfer in animals is commonly used to understand the role of genes in development, physiology and disease. This involves a primary step of screening for transgenes in the animal’s tissue using PCR, normally this involves DNA extraction from a biopsy, ear punch or toe tissue. This can cause pain and stress to the animal, as well as being a time consuming method. This application note describes a time-saving method for genotyping from hair samples, the collection of these will cause minimal discomfort to the animal.

Arktik Thermal Cycler

Thermo Fisher Scientific

Thermo Scientific Arktik Thermal Cycler is suitable for a wide range of applications from individual reactions to high-throughput projects. It offers flexibility with three different interchangeable blocks: 96-well block, 384-well block, and a dual block with two 48-well units. A broad temperature gradient in 96 and 384 blocks simplifies temperature optimization. Ease-of-use is the key feature of the Arktik Thermal Cycler’s user interface. The protocol is displayed graphically, and the programming is simple with intuitive menus. Highlights of the Arktik Thermal Cycler: Interchangeable blocks with excellent thermal precision Dual 48-well block for multi-user option Broad (up to 30°C) and accurate temperature gradient† Reliable and easy to use Low noise level Accepts virtually all standard PCR plastics †In 96- and 384-well blocks. Gradient feature not available in the U.S., Germany, and Japan. Flexibility with interchangeable blocks Interchangeable blocks allow the Arktik Thermal Cycler to be quickly and easily transformed from a 96-well to a 384-well PCR instrument. With the dual block, the instrument operates as two 48-well units, enabling two independent protocols to be performed simultaneously. The blocks feature a patent pending heat sink design that improves the speed and thermal uniformity of the system. Broad and accurate temperature gradient speeds up protocol optimization* Tedious temperature optimization can be simplified to a single experiment with the Arktik Thermal Cycler’s advanced gradient feature. The gradient range is broad – up to 30°C – and accurate, allowing easy temperature optimization. Reliable and quiet instrument for demanding use State-of-the-art Peltier technology employed in heating and cooling of the blocks ensures that the instrument maintains its high level of performance in demanding use. An overtightening protection system in the heated lid simplifies setting the correct pressure for PCR vessels used. The low noise level of the instrument is an additional benefit when the instrument is placed in the laboratory working space.

(1)

Phire Animal Tissue Direct PCR Kit

Thermo Fisher Scientific

Thermo Scientific Phire Animal Tissue Direct PCR Kit has been developed for amplification of DNA directly from a wide variety of animal tissues including mice, fish, birds and insects. The Phire® Animal Tissue Direct PCR Kit employs Phire Hot Start II DNA Polymerase, a specially engineered enzyme with a DNA-binding domain that enhances the processivity of the polymerase. Phire Hot Start II DNA Polymerase also exhibits extremely high resistance to many PCR inhibitors found in animal tissues.The kit contains all the necessary components for amplification of DNA directly from animal tissues: optimized reagents, sampling tools, and DNARelease Additive, which can be used to improve the release of DNA from animal tissues. In addition, the kit includes detailed instructions and control primers that can be used with numerous animal species.Phire Animal Tissue Direct PCR Kit Features & Benefits: No need for time-consuming and expensive DNA purification steps Very little sample material required Two simple protocols for various applications Convenient sampling tools included in the kit Phire Hot Start II DNA Polymerase provides high yields of specific product with short extension time (20 s/kb). Minimized pipetting with master mix format and loading dyes pre-added Extremely short PCR protocol times Tested with wide variety of sample types Robust hot start DNA polymerases guarantee high yields of specific product Rich composition of the kit - controls, water and DNA ladder included Phire Animal Tissue Direct PCR Kit Includes: Phire Hot Start II DNA Polymerase 2X Phire Animal Tissue PCR Buffer (includes dNTPs and MgCl2) Universal control primer mix Dilution Buffer DNARelease Additive Gel loading dye Harris Uni-Core™ 0.50mm Harris Cutting Mat™

(0)

Links

Tags

Sample PreparationSample preparation can improve the quality and speed of separation techniques. Products to assist sample preparation include filtration equipment, evaporators, membranes and sieves.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.Data AnalysisData analysis hardware and software is available to make data processing straight-forward yet powerful. Data software can be used for math and stats, technical graphing and image analysis. In addition, software is available for specific data analysis of electrophoresis, densitometry, ELISA and DNA sequencing.Sample ManagementSample management systems include sample storage devices such as freezers and plate storers, sample environment enclosures and sample organization, retrieval and sorter systems. Useful system features include high-throughput, automation, robotic arms, automated liquid handling and associated database systems. Accessories in sample management include barcode scanners, heat sealers and tubes.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.RNA InterferenceRNA interference (RNAi) uses siRNA or miRNA for transcriptional silencing, gene knockdown and regulation of gene expression. RNAi requires chemical synthesis, introduction of DNA vectors into cells, an assay of RNAi effects and RNAi quantification or analysis. Consider target sequence selection, reagent preparation, controls, high specificity and effectiveness and low non-specific gene knockdown.Animal ModelsThe use of non-human animals in experiments or behavorial observations. The research is conducted inside universities, medical schools, pharmaceutical companies, farms, defence establishments, and commercial facilities that provide animal-testing services to industry. It includes pure research such as genetics, developmental biology, behavioral studies, as well as applied research such as pharmaceutical testing in pre-clinical, before human, studies. RegulatoryPharmaceutical regulations impact on all areas of drug development, manufacture and control and supply. Services are provided from the early stages of drug development and clinical trials through to dossier submission, approval and marketing. Regulatory also includes submission of analytical studies from the pharmaceutical product and safety. PCR AutomationPCR automation describes the process of using a thermocycler to complete the polymerase chain reaction (PCR) process quickly and efficiently.GenotypingSample HandlingProper and careful handling of samples is important in scientific studies to ensure the integrity of specimens. This will prevent deterioration and cross contamination, and improve the accuracy of sample tracking procedures.Sample ProcessingSample processing is required to prepare samples prior to analysis. Technologies used for sample processing include homogenizers, grinders, mills and mixers.
Direct PCR on Hair: A New Animal-Friendly Genotyping Method