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. 2010 Jan 15;6(1):e1000799. doi: 10.1371/journal.pgen.1000799

Figure 3. Physical evidence for irradiation-induced ISDra2 excision.

Figure 3

Strain GY13120 (tetAΩISDra2-113 expressing tnpA in trans) received 5 kGy of γ–irradiation and aliquots were taken to isolate genomic DNA used to prepare DNA agarose plugs and as template for PCR analysis. The time following irradiation is shown in hours above each lane. (A) Schematic representation of IS excision products. The excision products are shown together with the position of primers used in PCR analysis. Note that the transposon circle including the transposon junction is single stranded. (B) Kinetics of double-strand-break repair. DNA agarose plugs were digested with NotI prior to PFGE analyses and loaded onto a 1.06% agarose gel. L: λ Ladder. (C) Kinetics of donor joint appearance. PCR reactions were performed with primer pair P1 and P2 and loaded onto a 1% agarose gel; L: MassRuler DNA Ladder. (D) Kinetics of IS circle junction appearance. PCR reactions were performed with primer pair P3 and P4 and loaded onto a 2% agarose gel; L: O'GeneRuler 100 bp DNA Ladder.