Accurate and reproducible quantification of DNA is essential to achieving consistent NGS sequencing results. It is important to measure both the total quantity of DNA and the distribution of fragment sizes to achieve an optimal cluster density on the Illumina flow cell. Both factors impact data quality and total sequencing output. Among the common methods of DNA quantification, qPCR has the highest sensitivity.
However, qPCR setup is time intensive, and results are highly sensitive to small differences in pipetting. Automating NGS quantification provides a streamlined approach to getting reproducible, high-precision results while minimizing hands-on time.
This application note from Opentrons describes studies performed to evaluate two methods of qPCR normalization during the NGS workflow on the Opentrons OT-2 automated liquid handling platform, demonstrating the capabilities of the platform in automating NGS quantification and normalization in preparation for Illumina sequencing.
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