PIPETMAX®: Automation of the Illumina® Nextera® XT DNA Library Preparation Kit

The Illumina® Nextera® XT DNA Library Preparation Kit workflow was automated on PIPETMAX®. For comparison, 12 technical replicate libraries were prepared side by side, using manual liquid handling or automated on PIPETMAX. The 24 libraries were pooled and sequenced on an Illumina MiSeq® sequencing system. Both library preparation methods generated high quality data with >95% mapped reads and optimal quality scores. Variance was lower for libraries generated with PIPETMAX, consistent with the reproducible performance of this automated pipetting system. Automation of the Nextera XT System workflow with PIPETMAX allows libraries to be prepared in parallel and provides walk-away time, freeing up the researcher to carry out other tasks. More importantly, this application demonstrates the reliability and process control that is important for complex workflows, such as next generation sequencing. INTRODUCTION The Illumina® Nextera® XT DNA Library Preparation Kit1 is designed to rapidly prepare sequencing-ready libraries of plasmids or small genomes for next-generation sequencing (NGS), also referred to as massively parallel sequencing or deep sequencing.2 To automate this workflow, five automated scripts were developed for PIPETMAX3,4 (Figure 1). These scripts, accessed via Figure 1 PIPETMAX® is a benchtop, automated liquid handler equipped with motorized, multichannel, air displacement pipettes.