After ExoIII digestion, the resulting single-stranded regions are digested with S1 nuclease, and the ends are repaired and ligated with T4 DNA polymerase and ligase. Pooling samples from several different ExoIII digestion time points before subsequent S1 treatment generates a good distribution of deletion clones following electroporation. Deletion clones are sized and sequenced using vector specific forward and reverse primers. Funding for the Borrelia work was supported by grants from the NIH . Developed by Jack McNulty.
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