(1120-A) Efficient Automated Extraction of Plasmid DNA in Medium Throughput

Abstract: The isolation of plasmid DNA is an essential molecular biology technique for many downstream applications. The multi-step protocol is simple, but labor-intensive and error prone. The use of Promega MagneSil® Paramagnetic Particles for lysate purification and DNA capture bypasses the need for centrifugation or a vacuum manifold, allowing DNA purification to be fully automated with the Promega Wizard MagneSil Tfx™ System on the Analytik Jena Liquid Handler CyBio FeliX.
Escherichia coli JM109 bacteria, were transformed with pGL4.50 and cultured in Terrific Broth (TB) or Luria Broth (LB) medium. The subsequent extraction was performed on the CyBio FeliX. Spectrophotometry and gel electrophoresis were used to determine DNA concentrations, absorbance ratio and plasmid size.
The TB culture yielded higher plasmid amounts, which can be attributed to the higher culture density. The results show that the automated procedure significantly reduced processing time and minimized variability while maintaining high quality and purity of plasmid DNA. This study highlights the effectiveness of the Promega MagneSil Tfx™ System and Analytik Jena CyBio FeliX in the simple, rapid, and reliable isolation of transfection-quality plasmid DNA in a 96-well high-throughput format.