Plasmid Isolation
1. Single colonies from each transformation were chosen and broth cultures were grown
in 1.5 ml of LBkan75 by shaking overnight at 37°C. These cultures were arrayed in a 96
well deep-well microplate provided with the QIAprep 96 turbo Miniprep Kit (QIAGEN
Inc., Valencia, CA).
2. Plasmid DNA was purified using QIAGEN’s QIAprep 96 Turbo Miniprep Kit or
QIAprep Spin Miniprep Kit. (link E2-QIAprep_Miniprep_Handbook.pdf)
3. Briefly, bacterial cells were harvested by centrifugation from a 24 h culture of E. coli
DH5α cells, lysed using a modified alkaline lysis procedure, and the DNA was bound to a
silica-gel-membrane. DNA was washed and eluted using 80 µl of sterile distilled H2O.