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S-phase Increases CRISPR/Cas9 and ssODN Directed Editing.

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posted on 08.06.2015 by Pawel Bialk, Natalia Rivera-Torres, Bryan Strouse, Eric B. Kmiec

HCT116-19 cells were seeded at 2.5x106 cells in a 100mm dish and synchronized for 24 hours with 6uM aphidicolin then released for 4 hours. Synchronized and unsynchronized cells were electroporated at a concentration of 5x105 cells/100ul with CRISPR/Cas9 and 72NT ssODN under the standard reaction conditions. Following electroporation, cells were seeded in 6-well plates and allowed to recover for 48 hours before flow cytometry analysis was carried out. Correction efficiency (%) was determined by the number of viable eGFP+ cells. Each sample set was performed in duplicate and ± represent calculated standard deviation per sample. The unsynchronized data is the same shown in Fig 4. Statistical analysis was performed using two-sample unequal variance students T-test distribution to compare the value of correction efficiency between synchronized and un-synchronized cells when treated with CRISPR/Cas9.


S-phase Increases CRISPR/Cas9 and ssODN Directed Editing.