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The Salmonella enterica (S. enterica) subsp. enterica, strain 14028s (serovar Typhimurium) targeted single-gene deletion (SGD) mutant library contains a total of 3,773 individual genes deleted simultaneously across two collections of mutants differentiated by kanamycin or chloramphenicol resistance. The kanamycin-resistant mutant collection contains 3,517 mutants distributed among eleven 96-well plates. In
these mutants, a single gene is replaced by a cassette conferring the kanamycin resistance gene, and includes 9 double mutants that contain both kanamycin and chloramphenicol cassettes. Deletions were confirmed by the depositor.
Genes were targeted for deletion by primers designed to preserve the first and last 30 bases of each deleted gene. Gene replacement followed a modified Lambda-Red technique, with an added T7 RNA
polymerase promoter positioned in plasmid pCLF4 to generate a gene-specific transcript from the Salmonella genome directly downstream of each mutant.
Plate orientation and viability were confirmed for NR-29409.
Each inoculated well of the 96-well plate
contains approximately 50 µL of culture in Luria Bertani (LB) broth containing 60
µg/mL kanamycin supplemented with 10% glycerol.
Production in the 96-well format has increased risk of cross-contamination between adjacent wells. Individual clones should be purified (e.g. single colony isolation and purification using good microbiological practices) and sequence-verified prior to use. BEI Resources does not confirm or validate individual mutants provided by the contributor.
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biological materials provided by BEI Resources, please select this link: Compliance Requirements.
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