Wild type C. burnetii NMII, with cbu0533 deleted
A total of 1 ug of the DNA sample was used as the input for the Illumina TruSeq DNA PCR-Free kit. Fifty-five microliters of each sample were added to a Covaris microTUBE-50 AFA Fiber Screw-Cap tube, and the samples fragmented in the Covaris M220 focused ultrasonicator using the standard conditions for a 350 bp insert size (10% duty cycle, 50 watts peak power, 200 cycles per burst, 65 seconds). The samples were size selected using sample purification beads, and libraries generated using an Illumina TruSeq DNA PCR-free kit and the manufacturer recommended protocol (Illumina Document # 1000000039279 v00). Libraries were analyzed on an Agilent Bioanalyzer (Figure 1) and quantified using the Roche Kapa Library qPCR kit (Table 2). The individual libraries were diluted to 2 nM and 3 ul of each pooled. The library pool was denatured and diluted to a 9 pM stock and paired-end 2 x 150 cycle sequencing carried out on the MiSeq using a Micro V2 flow cell and 300 cycle chemistry.