Fluorescence Activated Cell Sorting generates higher quality independent clones than manual derivation.

<p>Modified pluripotent scorecard assay was performed on manually and FACS derived clones to demonstrate (<b>A</b>) activation of endogenous gene expression and (<b>B</b>) silencing of gene expression and presence of unreprogrammed and transformed fibroblasts CD13^POS in manually derived clones. (<b>C</b>) Three sorted and three picked lines from patient 1023 were used to compare the ability of both methods to generate independent clones. 10 μ of genomic DNA were cut overnight with BglII and submitted to Southern blotting. The HUES line HES2 was used as a positive control for endogenous KLF4/OCT4, and as a negative control for transgene insertions. Samples were first blotted for KLF4, then stripped and reblotted for OCT4. Picked clones 1023 C and E are consistent with being the same clone by both KLF4 and OCT4 blotting. * indicated the predicted endogenous KLF4/OCT4 bands, and ** indicated a consistent band found in all samples blotted with OCT4.