Tn-seq of A. actinomycetemcomitans: mutant pool prior to growth in a selective environment
INSeq method: Purified DNA was sheared, amplified by linear PCR with a single biotinylated primer specific to the Tn10 transposon, captured with streptavidin-coupled magnetic beads, made double-stranded, digested with NotI (engineered into the capture primer sequence) to remove the biotin tag, and then, using the NEBNext kit, end-repaired, A-tailed, and ligated to Illumina adapters; two technical replicates were prepared using the INSeq method from a single freezer aliquot of the A. actinomycetemcomitans strain VT1169 Tn10 transposon mutant pool and sequenced on a paired-end 2 x 100 read HiSeq run; this data file was generated by concatenating the read 1 and read 2 fastq files from the two technical replicates and then filtering for the Tn10 inverted repeat sequence (present only in reads truly derived from the transposon)
SUB1333650
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