PacBio Sequel IIe 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 for the PacBio Sequel IIe system were created using the SMRTbell Express Template Prep Kit 2.0 (n=3) or SMRTbell Prep Kit 3.0 (n=3). Size selection was performed using 3.7X 35% Ampure SPRIselect (Beckman Coulter), which targeted the removal of fragments <3kb. Six libraries were sequenced on the Sequel IIe system (using 4060 ng library input) with a 8M SMRT Cell each.