Genome sequencing of synthetic Escherichia coli, C321.ĆA.709
gDNA prep. cell culture was processed with a Qiagen DNeasy Blood and Tissue (cat: 69504) to extract genomic DNA. Sequencing. 2.5 g of gDNA was sent to the Yale Center Genome Analysis for library prep. One micrograms of gDNA were sheared to an expected size of 500 bases with Covaris E210 in a covaris microtube. Post-shearing cleanup was done with SPRI magnetic beads. ÔWith BeadŐ fragment end repair was performed with End Repair enzyme at 20űC for 30 min and purified. ÔWith beadŐ A-base addition was performed with A-Tailing enzyme at 30_C for 30 min and purified. Samples were barcoded with an adapter ligation mix. Ligation was purified. Samples were PCR enriched (26-l DNA; 30-l KAPA HiFi Mastermix; 2-ul 25-M PCR Primer MP1.0; 2-l 25-M barcode-specific primer). Samples were loaded onto a lane of an Illumina HiSeq 2000 for 76-base pairedend reads.