Multiplex Methods to Examine Epigentic Modifications

6 Jun 2018

This whitepaper explains how multiplex methods deep bisulfite sequencing and bisulfite PCR-Luminex can be used for DNA methylation analysis in research examining epigenetic modification.

Luminex® 200™Multiplexing Instrument

Luminex

The Luminex® 200™ detects up to 100 protein or nucleic acid analytes from a single aliquot of sample in a single reaction well, enabling you to generate more data while saving sample, time, and reagents. With more than 11,000 units sold globally to date, the Luminex 100/200 is the established standard in multiplexing instrumentation. The xMAP® multiplexing platform is composed of an instrument and associated reagents: The Luminex 200 instrument is based on the principles of flow cytometry, which integrates key xMAP detection components such as lasers, optics, fluidics, and high-speed digital signal processors. xMAP microspheres are a family of fluorescently dyed carboxylated polystyrene microspheres that act as both the surface for the solution phase assay and the spectral identifier that the instrument detects. The open architecture of xMAP Technology enables users to build custom multiplex assays or select from a broad menu commercially available kits. Instrument Details: Multiplex Capacity: Up to 100 non-magnetic; 80 magnetic Microtiter Plate: 96 well Throughput: ~40 min/96-well plate (up to 12,800 tests per hour) Dynamic Range: 3.5 logs Sensitivity: Detects a minimum of 1,000 fluorochromes of phycoerythrin (PE) per xMAP microsphere Optics: Lasers/APDs/PMTs Hardware: Flow Cytometry based Software: xPONENT® Software supports protocol-based data acquisition with robust data regression analysis Dimensions: All components combined (Analyzer, XY Platform and Sheath Delivery System) 26.5" W x 23.5" D x 12.5" H (67.3 cm W x 59.7 cm D x 31.8 cm H) Weight: All components combined (Analyzer, XY Platform and Sheath Delivery System) 113 lbs (49 kg) xMAP Reagent Compatibility: Magnetic and nonmagnetic microspheres

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FLEXMAP 3D® Multiplexing Instrument

Luminex

The FLEXMAP 3D® detects up to 500 protein or nucleic acid analytes from a single aliquot of sample in a single reaction well, enabling you to generate more data while saving sample, time, and reagents.  Fast read time, compatibility with both 96- and 384-well plates, and ease of integration with front-end automation systems make the FLEXMAP 3D the platform of choice for high-throughput bioassay applications. The xMAP® multiplexing platform is composed of an instrument and associated reagents: The FLEXMAP 3D® instrument is based on the principles of flow cytometry, which integrates key xMAP detection components such as lasers, optics, fluidics and high-speed digital signal processors. xMAP microspheres are a family of 500 fluorescently dyed carboxylated polystyrene microspheres that act as both the surface for the solution phase assay and the spectral identifier that the instrument detects. The open architecture of xMAP Technology enables users to build custom multiplex assays or select from a broad menu commercially available kits. Instrument Details: Multiplex Capacity: 500 Microtiter Plate: 96 well and 384 well Throughput: ~ 20 min/96-well plate (up to 144,000 tests/hour); ~ 75 mins/384-well plate (up to 153,600 tests/hour) Dynamic Range: 4.5 logs Sensitivity:  Detects a minimum of 500 fluorochromes of phycoerythrin (PE) per xMAP microsphere Optics: Lasers/APDs/PMTs Hardware: Flow Cytometry based with dual sample fluidics paths and rapid sampling rate Software: xPONENT® Software offers interfacing options for front-end automation platforms and Laboratory Information Systems (LIS) Dimensions: 23” W x 25.7” D x 18” H (58.4 cm W x 65.3 cm D x 45.7 cm H) Weight: Up to 91 kg (200 lbs), including the instrument, monitor arm, monitor, and accessories, such as the keyboard, mouse and barcode scanner xMAP Reagent Compatibility: Magnetic and nonmagnetic microspheres

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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.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.Epigenetic AnalysisEpigenetic analysis products are used to study heritable phenotype changes that do not alter the DNA sequence, such as DNA modifications affecting gene activity. Determine whether genes have been methylated with DNA methylation quantification kits or methylation microarrays. Assess epigenetic regulation with histone modification kits or ChIP-on-chip microarrays. Kits are also available for bisulfite conversion and amplification of your samples, as well as epigenomic library preparation kits for next-generation sequencing (NGS). Find the best epigenetic products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA MethylationDNA AmplificationDNA Amplification is a technique used to amplify a single or multiple copies of DNA or mRNA by orders of magnitude. The most common method employed is PCR, but other options exist that eliminate the need for thermocycling.PCR PurificationDNA TypingBiomarker DiscoveryClinical biomarkers refer to substances related to known medical conditions that can be accurately measured <i>in vitro</i>. Biomarkers can be used to diagnose presence of a disease and indicate disease severity. The discovery of new biomarkers is incredibly valuable in the field of diagnostics.