Automated NGS Library Prep: Ligation Efficiency Under Observation

摘要

As the number of samples for sequencing increases, there is a need for library prep automation that matches the throughput of the next generation sequencing (NGS) instruments while not requiring an investment equal to the sequencer itself. By utilizing the Gilson PIPETMAX 268, a novel NGS sample prep chemistry was automated to reduce the time to achieve the final prepared library, increasing the efficiency of the method and consistency of the resulting DNA library to be sequenced. The here presented sample prep chemistry combines typical end-repair and A-tailing steps into one master mix step with buffers directly compatible with downstream ligation steps, eliminating the need for multiple cleanup steps throughout the process. The chemistry has also been optimized to drive higher A-tailing efficiencies, which reduces chimera formation from blunt fragments and loss of fragments tailed with nucleotides other than the necessary "A." Further, these chemistries allow numerous DNA libraries to be prepared simultaneously, including incorporation of barcoded adapters for multiplex PCR and sequencing.