Before you begin
This guide describes the workflow for preparing Sequencing By Binding (SBB™) libraries for cluster generation on
the PacBio
®
Onso™ cluster generator system, as well as preparation for and sequencing of clustered flow cells on
the Onso system. Below are descriptions of the main consumables required for both clustering and sequencing
procedures.
Flow cell and Main Instrument Connector Assembly (MICA) configuration
Each clustering and sequencing run requires use of a single flow cell that comes pre-attached to two Main
Instrument Connector Assembly (MICA) units as shown below.
The (A) flow cell immobilizes DNA clusters for sequencing and interfaces with the instruments’ mechanical, fluidic
and optical systems. The flow cell interfaces with the fluidics systems via the (B) pre-attached MICA during
clustering and sequencing.
Flow cell configuration
Each flow cell contains two lanes for sequencing libraries, which experience opposing directional flow. Lane 1 (L01)
experiences flow of reagents from left to right, while lane 2 (L02) experiences flow from right to left.
(A) Ports on the left- and right-hand side of each flow cell allow for the flow of reagents into and out of the flow cell
via the attached MICA. (B) Pinholes on the wings of the flow cell align with metal pins on the cluster generator and
sequencing instrument for proper flow cell loading. The left side of the flow cell will have two pinholes, while the
right side contains one. A QR code (C) on the upper right corner contains flow cell information and indicates the
correct orientation when loading onto the instruments.
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