PacBio Revio sequencing of pooled human gut microbiome (Zymo D6323)
DNA was extracted using the ZymoBIOMICS DNA Miniprep Kit (D4300), which involved either mechanical (Vortex Genie 2) or enzymatic lysis. Fragment sizes were measured using a Femto Pulse (Agilent). Libraries were prepared for the Revio system using the SMRTbell Prep Kit 3.0 (n=2). Size selection was performed using 3.7X 35% Ampure SPRIselect (Beckman Coulter), which targeted the removal of fragments <3kb. Two libraries were sequenced on the Revio system (using 130140 ng library input) with a 25M SMRT Cell each.