Agilent Technologies 4150 TapeStation system User Manual

206 Agilent 4150 TapeStation System Manual
10 Troubleshooting
Troubleshooting RNA Applications
Table 16 RNA - Possible causes of unexpected migration profile.
Problem Likely Cause Explanation Solution
Additional bands are
present in the region of
the 18S and 28S peak
Incorrect heating step The 18S peak may have
partially reverted back
to its original,
non-denatured
conformation which
which results in
characteristic double
peaks and influences
RIN
e
calculation
Ensure the correct denaturation conditions
during the RNA sample preparation, according
to the assay Quick Guide. Samples should be
kept on ice after preparation, and run within 2
hours of denaturation (see “Heat
Denaturation” on page 99) (Table 3 on page 64)
Genomic DNA
contamination
Samples that are
contaminated with
genomic DNA contain
additional signals
migrating in and
beyond the region of
the 18S and 28S.
Occasionally this peak
can be mistaken for the
18S or 28S peak
Ensure that all peaks are annotated correctly
and correct if this is appropriate. Treat samples
with DNAse to eliminate the genomic DNA
peak then prepare a new run.
See the Agilent Information Center for more
information.
Contamination with
prokaryotic total RNA
Ribosomal RNA peaks,
namely 16S and 23S,
are smaller than
eukaryotic 18S and 28S
rRNA peaks and as
such can be seen in the
analysis. See the
Agilent Information
Center for more
information.
If not intended or expected ensure that sample
tissue is free of prokaryotic contaminations.
There may be a
contaminant peak that
is caused by dust or dirt
on the ScreenTape
device
Peaks caused by
contaminants appear
as sharp strong bands
which are often slanted
or uneven in the gel
image, and are usually
easily distinguishable
from sample peaks
Unalign the image using the Aligned button in
the TapeStation Analysis software. Contaminant
peaks can be unassigned by right-clicking on
either the gel image or electropherogram peak.
Rerun samples if needed
A bubble could be
located within the gel
matrix
A bubble in the gel
matrix will introduce
significant artifacts in
the data. Those bubbles
cannot be flicked away
Always inspect ScreenTape devices for bubbles
in the lanes. Lanes in question cannot be used
for analysis. Such bubbles can form after the
expiration date or upon wrong storage. See the
Agilent Information Center or “ScreenTape
Devices” on page 87 for more information

Agilent Technologies 4150 TapeStation system Specifications

General

Brand	Agilent Technologies
Model	4150 TapeStation system
Category	Laboratory Equipment
Language	English