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Gene, Transcription Start Site and Splice Site Identification Increased Through Use of Automated Preparative Gel Electrophoresis

Sage Science
9 Sept 2013

Gel purification of cDNA and RNA-Seq libraries is the gold standard for purification options but these methods are time consuming and so most protocols have standardized on bead or column purification methods. This scientific poster compares two systems for library purification steps in NGS. The Pippin Prep, a preparative gel electrophoresis system, is shown to offer several advantages over other protocols for this application including efficient removal of low molecular weight library contaminants (primer/adapter artifacts) and accurate size-selection of the library.

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Pippin Prep DNA Size Selection System

Sage Science

The Pippin Prep is a preparative electrophoresis platform for collection of size-selected DNA samples. The system automates DNA size selection using disposable, pre-cast agarose cassettes to extract fractions according to user-defined software input. Samples are electro-eluted from agarose and collected in buffer. The platform is used for the construction of next-gen sequencing libraries. The system collects fragments (depending on gel type) between 90 bp- 8kb. A higher quality of sample can be extracted, more reproducibly and with no cross-contamination, when compared to manual gel purification methods.

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ElectrophoresisElectrophoresis equipment is used to separate mixtures of protein, DNA or RNA, based on their electric charge, size and other physical characteristics, by passing them through a medium such as a polyacrylamide gel, an agarose gel or a capillary tube. Electrophoresis equipment includes horizontal and vertical gel electrophoresis chambers, isoelectric focusing systems, 2D electrophoresis and capillary electrophoresis instruments. Precast gels with a gradient can be used or gels can be hand cast. Samples are run alongside ladders or markers to identify the approximate size of a molecule. Separated proteins and nucleic acids in the gel can be stained or probed with fluorescent markers and then imaged and detected using gel documentation instruments, transilluminators, densitometers and scanners. Find the best electrophoresis equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.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.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.Library PreparationLibrary preparation is a critical step in sequencing workflows, where DNA or RNA samples are converted into libraries for high-throughput, next generation sequencing. This step ensures accurate results and minimizes biases. Explore library preparation kits in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.NGS AutomationNGS automation streamlines next-generation sequencing workflows, increasing throughput, accuracy, and reproducibility. Automated solutions enhance productivity in genomic research, clinical diagnostics, and personalized medicine. Browse peer-reviewed tools to compare products, read reviews, and get pricing.RNA-SeqRNA sequencing (RNA-seq) is a next-generation sequencing technique used to analyze the transcriptome, providing insights into gene expression and regulation. This method is essential for genomics, disease research, and personalized medicine. Explore RNA-seq tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Size Selection

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Gene, Transcription Start Site and Splice Site Identification Increased Through Use of Automated Preparative Gel Electrophoresis