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Comparison of methods to generate a pooled Enterococcus faecalis Tn library from arrayed library plates

ABSTRACT: Comparison of methods to generate a pooled Enterococcus faecalis Tn library from arrayed library plates

PROVIDER: PRJNA975209 | ENA |

REPOSITORIES: ENA

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Comparison of methods to generate a pooled Enterococcus faecalis Tn library from arrayed library plates

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Project description:Microarray Tracking of transposon mutants for a H. pylori mouse colonization screen described in Baldwin DN et al. Screen in NSH57 H. pylori strain background. Original 50,000 clone transposon library was plated and patched to make 25 pools of 48 clones. Clones were infected into 4-8 C57Bl/6 mice and stomach bacteria from at least two mice were harvested at 1 week or one month. Semi-random PCR was used to amplify and label the DNA next to the transposon insertion from the input (Cy3) and output pool (Cy5) genomic DNA for each array. Two arrays were done per mouse. One array labeled from the left side of transposon (primers S, 2C) and one array labeled from the right side of the transposon (primers N3, 2C). Transposon insertions were defined by spots with signal four standard deviations above background in both arrays. We also counted insertions where two adjacent gene spots (after arranging the data in genome order) gave signal from the two different sides of the transposon (but not both). A pathogenicity experiment design type is where an infective agent such as a bacterium, virus, protozoan, fungus etc. infects a host organism(s) and the infective agent is assayed. Keywords: pathogenicity_design

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