MAGNETIC BEAD BASED PLASMID DNA PURIFICATION SYSTEM Description The Clean Plasmid TR DNA Kit is based upon our proprietary magnetic beads based system to extract high quality and endo toxin free plasmid DNA. The system can easily be automated on many liquid handling workstations (e.g. Beckman, Hamilton, Tecan, Caliper, Perkin Elmer, Dynamic Devices, Agilent and Eppendorf) providing robustness and reproducibility. Plasmid yields may vary according to E.coli strain used, plasmid copy number and growth conditions. Typically an overnight culture in 1 mL LB medium provides 10 up to 15 pg of plasmid DNA for high-copy plasmids. The isolated plasmid DNA is endotoxin free and directly suitable for transfections, Sanger sequencing, restriction enzyme digestion as well as other molecular biology procedures. Bacteria are resuspended and then lysed using our CleanNA buffer system. Once the lysis process has been stopped, the samples are centrifuged and a cleared supernatant is transferred into a new processing plate. Optional, our lysate clearance beads can be used to clear the lysate from any cell debris. Plasmid DNA is then bound to our magnetic beads in presence of our binding buffer. The CleanNA particles are separated from solution using a magnetic device. After a few wash steps, the purified plasmid DNA is eluted from the beads using an elution buffer. • Sanger Sequencing • Restriction digestion Coenecoop 75 2741 PH Waddinxveen The Netherlands

1 CleanNA, Clean Plasmid TR DNA Kit, using the lysate clearance beads 2 CleanNA, Clean Plasmid TR DNA Kit, using centrifugation 3 Company Q, spin column method 4 Company O, spin column method 5 Company P, magnetic bead method Plasmid DNA was isolated from 0.8 mL cultures (LB medium) after a 24 hour incubation. Isolations were performed in triplo, according the manufacturers recommended protocols. Graph above is showing the average data from each kit. DNA yield (pg) has been determined using the Denovix DS-11 spectrophotometer. Endotoxin levels in isolated Plasmid DNA Plasmid DNA was isolated...