Advancing the quality control methodology,,,to assess isolated total RNA,,,and generated fragmented cRNA
at times the 18S and/or 28S bands of the total
RNA were not clearly visible on
the gels making the interpretation
difficult. When running fragmented
cRNA samples on MOPS gels it
was often difficult to distinguish
whether or not the samples were
allowed a sufficient amount of
time to fragment because of poor
resolution. Problems with MOPS
gels would lead to re-running of
the samples on new gels, therefore
wasting valuable time and money.
The samples that should have
failed would be caught by quality
control measures later on in the
process, but would ultimately cost
the company money on reagents
in the steps leading up to that.
The Agilent 2100 bioanalyzer combined
with the RNA 6000 Nano
LabChip Kit provides key advantages
to running MOPS gels. The
Agilent bioanalyzer data is easy to
interpret and the chip runs on the
bioanalyzer are less time consuming.
It can take up to an hour and
half to prepare the samples, run a
gel, stain the gel, and take a picture.
The whole Agilent process
(sample preparation to saving the
file) can be done in half that time.
There is no improper staining and
no shortened or lengthened run
times. Also, because each chip has
the capacity to run 12 samples, at
approximately 30 minutes per
chip, this makes the Agilent 2100
bioanalyzer a useful tool for high
throughput screenings, especially
with multiple bioanalyzers.
The goal was to find a new assay
to be used to evaluate the quality
of the total RNA and fragmented
cRNA. This was needed because
there was occasional inconsistency
with the use of MOPS gels. The
qualitative analyses should be as
definitive and dependable as possible.
Problems associated with
MOPS gels in a high throughput
environment may be due to user
error. Common mistakes seen in
the laboratory are: accidentally
piercing the gel while loading, not
loading enough of sample into
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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