Choosing the
right electrophoresis instruments and consumables for your applications
Electrophoresis
is a technique that has become indispensable in life science labs. As such
there are many types of devices that vary according to scale and application.
This guide focuses on questions to ask when evaluating the right
electrophoresis devices and components for your needs.
What type of
electrophoresis is right for your application?
There are a
number of different types of electrophoresis that are commonly used, and each
type is better for certain applications:
- Agarose gel
electrophoresis techniques are optimal for separations of large molecules such
as nucleic acids. Applications include DNA and RNA fragment size analysis.
- Polyacrylamide
gel electrophoresis (PAGE) has smaller, more uniform pores allowing for
separation of small polymers and proteins. Applications including size and
charge analysis of native or denatured proteins.
- Capillary
gel electrophoresis (CGE) is performed using micrometer or nanometer diameter
sized glass capillaries. CGE is used for fast separations of nucleic acids and
proteins including DNA sequencing and protein profiling.
- Capillary
zone electrophoresis (CZE) performs solutions-based separations and can be
coupled with additional techniques such as capillary isoelectric focusing
(cIEF). Applications include protein charge variant and purity analysis.
What
resolution is required for your application?
Different
sizes of nucleic acids and proteins will directly affect the type of gel and
the instrument specifications that are ideal for the process:
- Agarose gel electrophoresis is most frequently used for large nucleic acid sequences,
ranging from 50bp to 20,000bp
- Polyacrylamide
gel electrophoresis (PAGE) is generally used for proteins (200 Da to 200 kDA)
or smaller nucleic acid (5bp to 500bp) sequences
- Capillary gel electrophoresis is best suited for fast, multiplexed
separations of nucleotide sequences and proteins using small sample volumes.
The technique can be scaled down for compatibility with microfluidics.
What sample
throughput is desired?
The sample
throughput requirements of the laboratory can have an impact on the type of
system required.
- For
laboratories that are running few nucleic acid or protein samples, there are a
variety of single or dual tank systems designed for mini gels.
- For
laboratories running a larger number of samples, there are larger-scale agarose gels to run up to 80 samples and midi gels offering the ability to run up to 12
full sized protein gels simultaneously.
- Capillary
electrophoresis systems can vary sample type, volume, and throughput for a
given application. DNA and RNA fragment analysis can involve a lower number of
samples (<96). DNA sequencing may involve large scale, high throughput
sample analysis (>96 using multiple microplates).
What
reagents and other consumables are needed for electrophoresis operations?
There are a
number of reagents and needed to properly run your electrophoresis and analyze
your results, including:
- Molecular
weight ladders used to determine masses of proteins and nucleic acids
- Stains used
to visualize protein or nucleic acid bands on gels. These include ethidium
bromide and related markers for agarose gels, and Coomassie blue and a range of
alternative staining reagents used for protein gel visualization.
- Running
buffers are needed specific to type and application of electrophoresis
separations.
- Polymerization
reagents including acrylamide, reducing agents, and polymerization catalyst are
required for PAGE.
- Agarose in
powder form is used to set up agarose separations
- Electrolyte
solutions are necessary for isoelectric focusing and capillary electrophoresis
applications
- Precast gels
or capillary tube packs and cassettes are available for PAGE and CGE
applications.
What
additional hardware is required for your electrophoresis application?
Electrophoresis
equipment is sold as a package that includes the required instrument(s),
consumables and supplies, and data analysis software if necessary. These
components are also offered as stand-alone products.
Additional
hardware to consider includes:
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