Genome sequencing of Chlamydia pecorum 18-13680-18FL
A library was prepared from 100 ng of genomic DNA by fragmentation of the sample using the Ion XpressTM Plus Fragment Library Kit (Thermofisher catalog no. 4471269) and incubating for 7 min at 37C before magnetic bead purification using ethanol washes.  DNA was eluted in low TE buffer.  DNA was ligated and barcodes added at 25C for 15 min followed by 72C for 5 min.  The library was purified again using the magnetic bead method and products of approximately 450bp in length were recovered using the E-gel precast system.  Amplification of the library was achieved using the following conditions: 1  cycle of 95C for 5 min followed by 9  cycles of 95C for 15 s; 58C for 15 s and 70C for 60 s.  The amplified library was purified and diluted 1:100 before real-time PCR quantification on a ViiA 7 against Escherichia coli standards using the Ion Universal Library Quantitation kit (Thermofisher catalog no. A26217).