Agilent 2019 - 24
Automated Nucleic Acid Sample Quality Control in NGS Workflows
The second criterion for QC of incoming samples
is the calculation of total DNA abundance. A
minimum of 450 ng gDNA is required for DNA
library preparation. Samples were quantified in
triplicate using the Qubit system. The Qubit assay
is the established quantification method of the
High Throughput Sequencing Unit. The
concentration analysis of the 4200 TapeStation
was used as control to double check the Qubit
quantification.
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A
0
B
700
Total DNA amount (ng)
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DIN
Based on empirical evidence, the High Throughput
Sequencing Unit had established a DIN threshold
of 7 for their incoming gDNA samples. Figure 3A
shows the DNA integrity of all FFPE samples in this
study. DIN values ranged from DIN 1.3 to 6.2, and
are well below the QC threshold of DIN 7. Samples
with lower DIN require a special review, and are
only subjected to library preparation if there was
approval to proceed from the customer. Depending
on the quality, modified protocols can be used.
However, in these cases, successful library
preparation cannot be guaranteed.
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100
0
0
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Sample
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Figure 3. The incoming QC consisting of quality analysis was performed using the Agilent 4200 TapeStation system and the Agilent Genomic DNA
ScreenTape assay. A) Qualification of the 88 gDNA samples with the DNA Integrity number (DIN), the red line indicates the incoming QC criteria for DNA
quality (DIN ≥ 7). B) Quantification of the 88 gDNA samples. The total DNA amount was calculated based on the measured concentration and the volume.
The eight gDNA samples marked in red were not further processed. The red line indicates the incoming QC criteria for DNA quantity (≥ 450 ng).
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Agilent 2019
Table of Contents for the Digital Edition of Agilent 2019
Contents
Agilent 2019 - 1
Agilent 2019 - 2
Agilent 2019 - 3
Agilent 2019 - 4
Agilent 2019 - Contents
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