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 The
Agilent 2100 bioanalyzer is the industry standard for RNA sample QC and
has replaced labor-intensive gel electrophoresis for this application.
It is also rapidly replacing gel electrophoresis for DNA fragment analysis
and SDS-PAGE analysis of protein samples. A unique feature of the Agilent
2100 bioanalyzer is that it can be used for both electrophoretic separation
and flow cytometric analysis of cell fluorescence parameters. This versatility
makes the Agilent 2100 bioanalyzer an indispensable tool for the molecular
biologist and biochemist. The Agilent 2100 bioanalyzer is the most successful
microfluidics-based platform available commercially, offering solutions
for the analysis of DNA, RNA, proteins, and cells. It answers our questions
within 30 minutes, delivering automated, high-quality digital data.
Quantity and Quality
Due to the omnipresence of RNases and the instability of RNA, quality
checks and sample quantitation are essential steps before any RNA-dependent
application. Microfluidics technology makes it easier and faster to analyze
samples using just 1 µL of an RNA sample. With the Nano Series II
kits, as little as 25 to 500ng/µL of total RNA or 25 to 250ng/µL
of mRNA can be analyzed using this electrophoretic technology. However,
by using the Pico Series II kits, as little as 50 to 5000pg/µL of
total RNA or 250 to 5000pg/µL of mRNA can be analyzed. The 2100
bioanalyzer expert software generates an RNA Integrity Number (RIN), a
quantitative estimate, and will calculate ribosomal ratios of the total
RNA sample. The RIN can be used as a standard integrity measurement that
is independent of concentration, instrument, or analyst. On the other
hand, the percentage of ribosomal impurities is calculated for mRNA samples
using the expert software.
Agilent's DNA Series II kits, which include the DNA 1000,
7500, and 12000 Assays, require only 1μL of material per analysis and
automatically allow the sizing and quantifying of PCR fragments and restriction
digests accurately and reproducibly. Some of the most widely used techniques
in molecular biology are PCR and RT-PCR. While in some cases it is sufficient
to detect the presence or absence of a PCR product, in many cases the
quantitation of this product and detection of unspecific amplification
is critical. The Agilent 2100 bioanalyzer is the tool of choice for automated
sizing and quantifying of PCR and RT-PCR products in single or multiplex
mode with unprecedented accuracy. With the DNA Series II kits, as little
as 0.1 to 50ng/µL DNA can be analyzed using this microcapillary
technology. The DNA 1000 Assay accurately sizes fragments of 25 to 1000bp
in length, while the DNA 7500 and 12000 Assays accurately size fragments
100 to 7500bp and 100 to 12000bp respectively.
If you express recombinant proteins, purify proteins, and
perform stability studies or check antibody qualites, then the Protein
80 and Protein 230 Series II kits, which require only 4µL per analysis,
provide a fast and easy way to analyze a wide range of samples using lab-on-a-chip
technology. When used with Agilent's 2100 bioanalyzer, each chip is capable
of providing size, purity, and concentration information for 10 different
protein samples at once. This lab-on-a-chip approach eliminates handling
SDS-PAGE slab gels, staining, or imaging steps. With the Protein Series
II kits, as little as 60 to 2000ng/μL can be analyzed using the Protein
80 chip or 15 to 2000ng/μL using the Protein 230 chip. These chips
accurately size proteins of five to 80kDa or 14 to 230kDa respectively.
The flow cytometry chip set for the Agilent 2100 bioanalyzer
allows scientists working with cells to perform simple flow cytometry
assays. Each chip acquires cell fluorescence data at two excitation wavelengths
(470 nm/630nm) and two emission wavelengths (525nm/680nm) from up to six
different cell samples. As little as 20,000 cells per sample are required
to detect fluorescence signals and analyze specific subpopulations. A
recommended cell density of 2.0x106/mL, 10µL, which contains
approximately 20,000 prestained cells, is loaded onto the chip and the
fluorescence intensities in two channels are measured for about 750 single
cells per sample.
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