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 res...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.展开更多
文摘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.