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Validation of a Magnetic Bead Mixer on a NGS Library System

Validation of a Magnetic Bead Mixer on a NGS Library System

by Sikander Gill, Rajwant Gill, Nick Bandy and Dong Liang

bead-vortexerReduced costs and higher throughput have made next-generation sequencing (NGS) more accessible to users in such diverse fields as diagnostics, infectious disease, food safety and public health. Increased capacity has created a challenge: establishment of an automated library preparation workflow to enable reliable and robust sequencing. Magnetic beads play a significant role in many steps of NGS library preparation, highlighting the need for their uniform distribution. Here we demonstrate an efficient bead mixer placed on the deck of a robotic workstation to keep the beads uniformly suspended during distribution. Beads were mixed by the on-deck magnetic bead vortexer and distributed to a 96-well plate containing 1xPBS for measurement by spectrophotometer. It was observed that vortexing during distribution gave very uniform distribution of magnetic beads across all wells; correlations of libraries prepared by the VERSA automated workstation were very highly correlated with libraries prepared manually (Pearson’s r2 > 0.990). The magnetic bead vortexer provides highly consistent bead distribution during library preparation representing a significant improvement to current automated protocols.

Figure 2- Standard curve of the magnetic bead suspension prepared by manual suspension
table 2- Purified amplified library concentrations for positive and negative controls from the manual and automated procedures within the acceptable limits

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