Bacterial cultures grown for 24 hours in LB broth were used in respective culture amounts. DH5α cells transformed with pGEM vector. O.D. calculated using Thermo’s NanoDrop 2000c.
E.Z.N.A.® Plasmid Midi Kit offers a novel and reliable method of isolating up to 200 µg of plasmid DNA in less than 60 minutes. This kit combines HiBind® membrane technology with the time-tested consistency of alkaline SDS lysis of bacterial cells to deliver high-quality plasmid DNA. The DNase/RNase Free HiBind® DNA Midi Columns operate by eliminating time consuming phenol-chloroform extractions and alcohol precipitations. Plasmid purification using the E.Z.N.A.® Plasmid Midi Kit follows a simple bind-wash-elute procedure thereby making it possible for multiple samples to be processed. Although yields vary according to plasmid copy number, E.coli strain, and conditions of growth, 50 mL of an overnight culture in LB medium will typically produce 200 µg of high-copy number plasmid DNA. When working with low-copy number plasmids, one can increase the starting culture volume up to 100 mL. High-quality DNA is ready for immediate use in routine molecular biology applications such as automated fluorescent sequencing, restriction enzyme digestion, and transfection screening.