| Design Description | An IonTorrent PGM (Life Technologies Inc., Carlsbad, CA) used for sequencing after extracting DNA from stationary phase cells using standard techniques. DNA was digested and sequencing adaptors ligated using an Ion Xpress Plus Fragment Library Kit (Life Technologies Inc., Burlington, ON) according to the manufacturer’s instructions. Adaptor-ligated DNA was size selected to a target length of 480 bp on a 2% E-Gel SizeSelect agarose gel, and Agencourt MAPure XP beads (Beckman Coulter, Mississauga, ON) were used for purification steps. An Ion Library Quantitation Kit was used to calculate the library dilution factor prior to amplification and enrichment with an Ion PGM Template OT2 400 kit on an Ion OneTouch 2 system. An Ion Sphere Quality Control Kit was used to quantify the percent enriched Ion Sphere Particles prior to sequencing with an Ion PGM 400 Sequencing Kit. |
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