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January 15, 2020

An Engineered DNA Ligase for Efficient Conversion of Input DNA During NGS Library Preparation

Authors: Matt Miller, Scott Baskerville, Nikki Dellas, David Elgart, Sandy Gomes, Vesna Mitchell, Jonathan Penfield, Judy Viduya, and Jonathan Vroom

Next-generation sequencing technologies have been applied in the field of molecular diagnostics, where they promise the potential for high specificity and sensitivity for detecting markers of disease. Widespread adoption of this evolving technology depends partly on improvement of sample preparation workflows. Sample preparation converts input sample DNA (e.g., cell-free DNA) into sequencing-capable library fragments by the appropriate attachment of adapter molecules to both ends of the input molecules. In this multi-step process, adapter ligation via the use of T4 DNA ligase is the limiting step in complete conversion of sample to ligated library fragments. Inefficiency at this step, particularly with low-concentration DNA samples, currently limits the sensitivity of NGS-based molecular diagnostics.

 

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