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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Article
Implementation of Automated Sample Quality Control in Whole Exome Sequencing
Author(s)
Elisa Viering1, Jana Molitor1, Leslie Friedmann1, Jana Petersen2, Marie Beckhaus2 and Stephan Wolf2
Full-Text PDF
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DOI:10.17265/1934-7391/2017.06.001
Affiliation(s)
1. Agilent Technologies, Inc., Waldbronn 76337, Germany
2. DKFZ Genomics and Proteomics Core Facility, High Throughput Sequencing Unit, Heidelberg 69120, Germany
ABSTRACT
Aims: High-quality DNA as
input material for a NGS (next-generation sequencing) workflow is essential for
the successful preparation of a DNA library. Additionally, DNA quality has a
strong impact on sequencing results. Therefore, it is important to include QC (quality
control) steps to assess size, concentration, molarity, and integrity of the
DNA during the workflow. Material and Methods: The WES (Whole Exome
Sequencing) workflow at the
Genomics and Proteomics Core Facility of DKFZ (the German Cancer Research Center) was performed with several QC steps: QC of the input material, QC of
intermediate products during library preparation, and QC of final libraries.
The Agilent 4200 TapeStation system, which offers automated sample processing,
was used to evaluate quantity, size, molarity, and integrity of the samples. Key Findings: The Agilent Genomic
DNA ScreenTape assay offers an unbiased genomic DNA integrity assessment, which
enables protocol adaption for optimized library preparation, for example,
selection of a suitable shearing protocol. Additionally, QC steps during
library preparation such as evaluation of library size, concentration, and
molarity ensure maximal sequencing output. Significance: The automated and fast high-throughput analysis of genomic
DNA with the 4200 TapeStation system helps to save labor time and costs.
Additionally, the easy-to-use system can be integrated as a QC tool into the
NGS workflow to ensure successful library preparation. QC steps enable the
confirmation of suitable library size and concentration for the workflow.
KEYWORDS
NGS, FFPE (Formalin-fixed Paraffin-embedded), WES, DIN (DNA Integrity Number), QC, 4200 TapeStation system.
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