Summary of mycobacteriophage mutants generated with BRED.

1Mutations generated were gene deletions, insertion of C-terminal His6 tags, nonsense mutations, or replacement of phage gene with gfp (Replace).2Plaques recovered from electroporation were analyzed by PCR as indicated.3Individual plaques were screened after re-plating of an initial mixed plaque. Values where plaques were recovered and tested from more than one mixed plaque are separated by a semicolon and combined for average percentage.4Plaques were screened by DADA-PCR.5A total of 195 individual plaques from ten initial mixed plaques were screened, three of which contained no mutants.6Plaques were screened by restriction digestion.7Plaques were screened genetically by complementation.8Plaques were screened with an upstream primer complementary to the tag sequence.9Substrate has 100 bp homology flanking BPs gene 54.10Primer is complementary to gfp.11BRED was performed in M. smegmatis mc2155��pJV62 cells, which express Che9c gp6120.12Plaques were screened by MAMA-PCR.