Agilent Technologies 4150 TapeStation system User Manual

Agilent 4150 TapeStation System Manual 205
Troubleshooting
10
Troubleshooting RNA Applications
Incorrect or Missing Peak Annotation
Unexpected Migration Profile
There are a number of issues which can affect migration and sample peak
profile. If the issues below do not describe your analysis file, please contact
your support representative.
Table 15 RNA - Possible causes of incorrect or missing peak annotation in order of probability.
Root cause Explanation Solution
Peak is not a
ribosomal RNA or
Lower Marker peak
Only specific RNA peaks (16, 18, 23, 28S) and
Lower marker peak will be annotated by the
software
Manually add any important peaks by
right-clicking on either the gel or
electropherogram and selecting add other peak.
See the Agilent Information Center for more
information
Extremely degraded
sample
The TapeStation Analysis software will only
recognize ribosomal RNA sample peaks at
expected positions. If no distinct peak is found
at that position, no peak will be assigned
The TapeStation Analysis software can
occasionally fail to recognize peaks extremely
degraded RNA samples. Ensure that Good
Measurement Practices are followed as
detailed in “Heat Denaturation” on page 99;
then prepare a new run (“Good Measurement
Practices” on page 81)
Incorrect heating
step
The 18S peak may have partially reverted back
to its original, non-denatured conformation
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 (Tab le 3 on page 64)
Genomic DNA
contamination
Samples that are contaminated with genomic
DNA contain a third peak which migrates in 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.
Treat samples with DNAse to eliminate the
genomic DNA peak; then prepare a new run

Agilent Technologies 4150 TapeStation system Specifications

General

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