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High Throughput Plasmid DNA Preparation Using A Magnetic Bead Kit On An Automated Workstation

James Batchelor, Lynn Jordan, and Ed Alderman
Applied Science and Technology Department, Caliper Life Sciences, Inc., Hopkinton, MA 01748

Karen Heins and Olaf Stelling
Agencourt Bioscience Corporation, 100 Cummings Center, Suite 107J, Beverly, MA 01915

Introduction

The goal for plasmid purification is isolation of nucleic acid from cellular material. This can be accomplished through the use of the SPRI-based paramagnetic microparticles and an external ring magnet plate. These microparticles are encapsulated with a ligand that binds nucleic acids. Once bound, the microparticles are drawn toward the wall of the microplate by a separate, external ring magnet plate. Waste material can be withdrawn from the well leaving the particles (and nucleic acid) behind. The particles are washed and the nucleic acid eluted from the microparticles.

This technology lends itself to high throughput environments. Centrifuges and vacuum pumps with filters are not needed to separate the nucleic acid from the waste.

Materials and Methods

Instrumentation:

Liquid transfers were performed with a Sciclone ALH 3000 fitted with a High Volume Head and using 100 μL and 200 μL Automation Certified Pipette Tips. A deck mounted Microplate Shaker (CaliperLS), was used for bead resuspension.

Microplates:

- 96 well 1.1 mL Culture Block (Marsh #DW9611)
- 96 well round bottom microplate (Corning #3765)
- 96 well Ring Magnet Plate (Agencourt P/N 000219)

Reagents:

- CosMCPrep Kit (Agencourt P/N 001046)
- Contains Resuspension/Elution, Lysis, Neutralization Buffer and Microparticles in binding buffer
- Isopropanol - 70% EtOH

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