Gradient Gel Electrophoresis
Denaturing Gradient Gel Electrophoresis (DGGE) is an important technique used in the search for mutations and DNA polymorphisms critical in genetic disorders and cancers, and to understand genetic diversity among species. The new omniPAGE VS20WAVE-DGGE is a flexible system that can be fully customised to perform several different mutation detection techniques. The VS20WAVE-DGGE is a versatile single-unit solution for different single-base pair mutation detection methods. The newly designed VS20WAVE-DGGE is a complete system for DNA mutation analysis.
Using the innovative vertical screw-clamp technology of the new VS20-WAVE system, the VS20WAVE-DGGE is fully equipped with temperature control unit, stirrer, and gradient mixer and casting accessories to perform specific mutation analysis applications. A powerful microprocessor-controlled PID temperature control unit enables various mutation detection techniques to be undertaken between ambient temperature and 70C, while the simple fourscrew design of the WAVE insert accelerates set up of denaturing PAGE gels. Accordingly, the new VS20-DGGE can be used to screen single-base pair changes in the following applications:
The flexibility provided by the modular design of the VS20WAVE-DGGE and its wide range of accessories enables laboratories to switch quickly and easily between different mutation detection techniques, thereby maximising throughput and screening efficiency. A maximum 96-sample throughput allows detection of as many mutations within a couple hours, alleviating many of the bottlenecks associated with screening for DNA mutations. -
GM100 Gradient Mixer ensures efficient gradient formation
The GM100 gradient mixer is supplied as standard to ensure efficient gradient formation by mixing and delivering high- and low-density denaturant solutions. A flat-base design and support handle allows the GM100 to be secured to a retort stand, enabling it to be easily mounted on a magnetic stirring plate (e.g. CSL-STIR), while the mixing chamber can accommodate a magnetic stirrer to form a linear gradient. The MU-D01 peristaltic pump is also recommended for delivery of linear and reproducible gradient gels.
Innovative casting and set-up mechanism
The VS20WAVE-DGGE utilises novel vertical screw clamp technology to assemble two vertical gels. This reduces the number of screws required for set up making casting assembly faster, while a built-in inner buffer chamber within the PAGE insert allows set-up to be completed without the inclusion of heavy top tanks or buffer chambers. A dual purpose PAGE insert eliminates the need for plate transfer, and is used with a cam casting base to guarantee efficient leak free casting.
Utilises the same combs and accessories as standard VS20 systems
The VS20WAVE-DGGE can use all of the combs, glass plates and accessories of existing VS20 units, providing full flexibility. Two 1mm 24-sample combs are supplied as standard while optional 48-sample combs allow screening to take place directly from 96-well thermal cycler blocks after PCR amplification.
Parallel DGGE Overview
Parallel DGGE is based on the application of a
denaturant concentration gradient - increasing
in the direction of electrophoresis within a
polyacrylamide gel - and uniform temperature to
induce the partial unwinding or melting of
double-stranded DNA by distinct domains in a
sequence-specific manner. Melting breaks the
hydrogen bonds holding together each base pair
within a DNA sequence: with domains rich in GC
base pairs melting at higher temperatures
than those with high A-T base-pair content.
Parallel DGGE is based on the application of a denaturant concentration gradient - increasing in the direction of electrophoresis within a polyacrylamide gel - and uniform temperature to induce the partial unwinding or melting of double-stranded DNA by distinct domains in a sequence-specific manner. Melting breaks the hydrogen bonds holding together each base pair within a DNA sequence: with domains rich in GC base pairs melting at higher temperatures than those with high A-T base-pair content.
Incorporating sequence high in G-C content into the DNA target sequence by PCR® amplification (GC-clamping) prevents the DNA target with the lowest melting domain from melting completely at its predetermined Tm, which is maintained by the application of a constant temperature across the gel and within the buffer. Because both gel temperature and increasing denaturant concentration act to melt DNA in a sequenceand domain-specific manner, any point mutations amplified within a PCR® product will affect its Tm so that its subsequent mobility within a polyacrylamide gel differs from its wild type counterpart (see schematic diagram).
Consequently, the sensitivity of this technique in distinguishing individual DNA sequences, which might differ only by a single point mutation, has proven highly attractive to scientists screening for unknown mutations.
|Ordering Information||Buy Online|
|VS20WAVE-DGGE||Complete DGGE system with accessory kit and Casting Base 110Volts||$4200.00|
|Recommended power supply EV245 (replaces EV243)||$699.00|
|Combs||Click for combs|
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