FIG. 7.

Analyses of reinsertion events in the presence of puromycin selection. (A) Experimental procedure to analyze for reinsertion events. After the transfection of SB transposase, tTR and control clones were inoculated into 96-well culture plates with the indicated cell numbers. Excision-positive cell populations were expanded, genomic DNA was isolated, and LM-PCR analyses were performed to identify reinsertion sites. (B) Screening for excision events. Excision-positive wells derived from 100-cell/well groups theoretically contained one excision event, and insertion sites were determined by LM-PCR. M, molecular marker. (C) Flanking transposon genomic sequences. The original donor site is located on chromosome 2, and reinsertion chromosomes are shown.