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NR-42892??Salmonella enterica subsp. enterica, 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_160/161_Cm(Mutant Bacteria)|Salmonella enterica subsp. enterica|14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_160/161_Cm|-80°C or colder|M McClellandAcknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Salmonella enterica subsp. enterica, Strain 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_160/161_Cm, NR-42892."|Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.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 chloramphenicol-resistant mutant collection contains 3,376 mutants distributed among eleven 96-well plates, in which a single gene is replaced by a cassette conferring the chloramphenicol resistance gene, and includes 4 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
pCLF3 to generate a gene-specific transcript from the Salmonella genome directly downstream of each mutant.Note: The strain designation on the plate, strain CDC 6516-60, is incorrect. The correct strain designation is strain 14028s. S. enterica subsp. enterica, strain 14028s was originally known as strain 14028. A variant of the original strain with a rough colony morphology was designated 14028r and the original smooth strain was renamed 14028s. Strain 14028 is a descendent of strain CDC 6516-60, which was isolated from pools of hearts and livers of 4-week-old chickens. The complete genome of S. enterica subsp. enterica, strain 14028s (GenBank:
CP001363.1) and plasmid (GenBank:
CP001362.1) sequences are available.Each inoculated well of the 96-well plate contains approximately 50 ?L of culture in Luria Bertani (LB) broth containing 20 ?g/mL chloramphenicol 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.