High-Throughput, Transfection-Grade Plasmid Purification without Centrifugation Using Paramagnetic Particles

Part # PS359

Abstract

C. Leroy¹ , N. Bodet¹, B. Kiley ¹ , M. Blémont² , F. Bosco² , K. Kershner¹ and T. Schagat^1
1Global Services and Support, Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711;
²Innate Pharma, France.

Here we present protocols for high-throughput plasmid purifications using 96- or 24-deepwell plate formats on liquid handler platforms that overcome the challenges of: 1) centrifugation steps; 2) purifying enough plasmid for screening, transfection and library storage purposes; and 3) obtaining transfection-grade quality plasmids with little to no inhibitors after purification.

These protocols are fully automated with no hands-on intervention required during the purification process. In addition to standard miniprep scale (1.5ml input culture) we also demonstrate the efficiency of our purification protocols from large volume bacterial inputs (up to 30ml bacterial culture input) to maximize plasmid yields. Finally, our protocols contain a specific endotoxin removal step to obtain high-quality plasmids suitable for cell transfection. These plasmid purification protocols were developed to simplify the engineered antibody screening workflow.

Printed in USA.