Advancing the quality control methodology,,,to assess isolated total RNA,,,and generated fragmented cRNA
This Application Note describes how Gene Logic, Inc., has improved its
ability to analyze the quality of total RNA and fragmented cRNA by using
the Agilent 2100 bioanalyzer. The Eukaryote Total RNA Nano and Pico
assays are currently being utilized to examine the 28S/18S ratio in the
total RNA as well as detection of possible degradation. The mRNA Smear
Nano assay is used to confirm that a high percentage of the cRNA sample
is fragmented to lengths in the optimal range necessary to be hybridized
onto Affymetrix microarrays.
Gene Logic Inc. is a leading
provider of integrated genomics
information and bioinformatics
products and services related to
gene activity in human disease
and toxicity that enable global
pharmaceutical and biotechnology
companies to optimize the time,
risk and cost of drug discovery
and drug development. In order to
do this, the quality control methods
must ensure that only the
highest quality samples proceed
through our high-throughput
process, which in turn help to
reduce the downstream costs of
cDNA synthesis, In-Vitro Transcription
and, finally, hybridization
onto microarrays. For qualitative
purposes, many laboratories continue
to analyze their RNA samples
using MOPS gels with gel
electrophoresis. However, an
internet search of Microarray
facilities around the country
shows an increase in the usage of
the Agilent bioanalyzer.
When using gel electrophoresis to
examine the quality of samples,
pre-cast MOPS gels were used.
Every now and then a problem
with the gels was encountered,
either user related or gel related.
Some general problems occurring
were sample migration, mold
spots in the pre-cast gels and
staining difficulties. The key issue
associated with the RNA quality
control gel was that
Source:Page: All 1 2 3 4 5 6 Related biology technology :1
. Using the Agilent 2100 bioanalyzer for
quality control of protein samples prior to
. Is Z factor the best assessment for the quality of cellular assays delivering higher content?3
. Evaluation of RNA isolated from human tissues: quality control for surgical and postmortem samples4
. Quality control of antibodies using the
2100 bioanalyzer and the Protein 200
. Homogeneous, chemiluminescent kinase assay kits for assessing the functional activity of a broad range of serine-threonine and tyrosine kinases6
. Easy preparation of total RNA from cultured cells with TRIzol and
Eppendorf Phase Lock Gel
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Chaps Cell Extract Buffer can be used to lyse cells under nondenaturing conditions and is recommendedfor the preparation of cell lysates to be used with our caspase signaling pathway antibodies...Biology Products: