RBTnseq barcode amplicon data of Caulobacter infected with phage Circe
Samples were collected at 0 (uninfected), 1.5, 3, 4.5, and 7.5 hours post-infection. At each time point, 10 ml of culture for the 0, 1.5, 3, and 4.5 hour samples or 20 ml of culture for the 7.5 hour samples was removed, pelleted by centrifugation (12,000 x g, 3 min, 4C), washed once with 1 ml PYE, and stored as a cell pellet at 20C. Cell pellets were resuspended in 1020 l sterile water and used as template for PCR amplification of barcodes using Q5 polymerase (New England Biolabs) in 20 l reaction volumes. Each reaction contained 1X Q5 buffer, 1X GC enhancer, 0.8 U Q5 polymerase, 0.2 mM dNTPs, 0.5 M each of the universal forward primer (Barseq_P1) and a unique indexed reverse primer (Barseq_P2_ITxxx). Thermocycling conditions were: 98C for 4 min; 25 cycles of 98C for 30 s, 55C for 30 s, and 72C for 30 s; followed by 72C for 5 min and hold at 4C. PCR products were pooled and subjected to single-end 50-bp sequencing on an Illumina NovaSeq using TruSeq primers. This is time 0, replicate 2