...Follow this protocol to perform amplification of CRISPR pooled plasmid libraries in Escherichia coli ...Estimated time 2-3 hours) Transformation should be performed at the end of the day to ensure that growth times...shaking period, pool and gently mix the four tubes. Perform sequential 1:100 dilutions of the cells (add 10...10 µL of the pool to 990 µL LB then perform a second and then third 1:100 dilution). Plate 100 µL of each... recombined fraction but care must be taken to perform adequate NGS based QC to ensure no change in representation...to the pre-amplified stock. A last option is to perform a gel extraction of either the original sample ...of the DNA, but please note that NGS should be performed to ensure representation is maintained. Maxipreps...