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. 2014 May 1;9(5):e96483. doi: 10.1371/journal.pone.0096483

Figure 4. Gene editing of synchronized and released HCT116–19 cells using TALENs and ssODNs.

Figure 4

HCT116–19 cells were seeded at 2.5e6 cells in a 100 mm dish and synchronized for 24 hours with 6 uM aphidicolin then released for 4 hours before being electroporated at a concentration of 5e5 cells/100 ul with TALENs and the 72NT ODN under the standard reaction ratio TALEN:ssODN (2 µg/1.35 µg). TALEN amounts reflect the total TALEN plasmid added to each sample in equal portions. Following electroporation, cells were seeded in 6-well plates and allowed to recover for 48 hours and analyses took place on a Guava EasyCyte 5HT flow cytometer (see Materials and Methods). Correction efficiency (%) was determined by the number of viable eGFP positive cells divided by the total viable cells in the population. Each sample set was performed in duplicate and error bars represent standard error.